KR101939150B1 - Consensus prostate antigens nucleic acid molecule encoding the same and vaccine and uses comprising the same - Google Patents

Abstract

Common amino acid sequences of prostate antigens capable of destroying tolerance are provided herein in targeted species, including PSA, PSMA, STEAP and PSCA antigens. Also provided are vaccines which express one or more common amino acid sequences of the prostate antigens PSA, PSMA, STEAP and PSCA, as well as gene constructs / vectors and corresponding sequences. Also provided is a method of generating an autoimmune response to prostate cancer cells by administering one or more of the vaccine, protein and / or nucleic acid sequences provided herein.

Description

TECHNICAL FIELD [0001] The present invention relates to a common prostate antigen, a nucleic acid molecule encoding the same, a vaccine containing the same, and a vaccine comprising the same. [0002]

The present invention relates to a nucleic acid sequence encoding a consensus prostate protein and a fragment thereof; An improved method for inducing an immune response against prostate cancer cells, and an improved method for prophylactically and / or therapeutically immunizing an individual against prostate cancer.

Prostate cancer is an important therapeutic immune target. The occurrence of an immunotherapeutic approach is complex, that is, the immunogen needs to be generated so as to induce a strong immune response which preferably induces a CTL response.

Direct administration of nucleic acid sequences for vaccination against animal and human diseases has been studied and much effort has been focused on effective and efficient means of nucleic acid delivery to obtain the required expression of the desired antigens and immunogenicity and ultimately Get the success of this technology.

DNA vaccines have a number of conceptual advantages over more traditional vaccination methods, such as attenuated live viruses and recombinant protein-based vaccines. DNA vaccines are well tolerated in humans by preclinical studies that are safe, stable, readily generated, and show little evidence of plasmid integration [Martin, T., et al., Plasmid DNA malaria vaccine: the potential for genomic integration after intramuscular injection. Hum Gene Ther, 1999. 10 (5): p. 759-68; Nichols, W. W., et al., Potential DNA vaccine integration into host cell genome. Ann N Y Acad Sci, 1995. 772: p. 30-9]. In addition, because of the fact that the efficacy of the vaccine is not affected by pre-existing antibody titers to the vector, DNA vaccines are suitable for repeated administration [Chattergoon, M., J. Boyer, and D. Weiner, Genetic immunization: a new era in vaccines and immune therapeutics. FASEB J, 1997. 11 (10): p. 753-63]. However, one major hurdle for the clinical adoption of DNA vaccines has been the reduction in immunogenicity of the platform when migrated to larger animals [Liu, M.A. and J.B. Ulmer, human clinical trials of plasmid DNA vaccines. Adv Genet, 2005. 55: p. 25-40]. Recent technological advances such as codon optimization, RNA optimization and addition of immunoglobulin leader sequences in DNA vaccine immunogen genetics have improved the expression and immunogenicity of DNA vaccines [Andre, S., et al., Increased immune response elicited by DNA vaccination with a synthetic gp120 sequence with optimized codon usage. J Virol, 1998. 72 (2): p. 1497-503; Deml, L., et al., Multiple effects of codon usage on expression and immunogenicity of DNA candidate vaccines encoding the human immunodeficiency virus type 1 Gag protein. J Virol, 2001. 75 (22): p. 10991-1001; Laddy, D. J., et al., Immunogenicity of novel consensus-based DNA vaccines against avian influenza. Vaccine, 2007. 25 (16): p. 2984-9; Frelin, L., et al., Codon optimization and mRNA amplification effectively enhances the immunogenicity of the hepatitis C virus nonstructural 3 / 4A gene. Gene Ther, 2004. 11 (6): p. 522-33].

Recent technological advances including techniques such as electroporation in plasmid delivery systems have improved the expression and immunogenicity of DNA vaccines [Hirao, LA, et al., Intradermal / subcutaneous immunization by electroporation improves plasmid vaccine delivery and potency in pigs and rhesus macaques. Vaccine, 2008. 26 (3): p. 440-8; Luckay, A., et al., Effect of plasmid DNA vaccine design and in vivo electroporation on the resulting vaccine-specific immune responses in rhesus macaques. J Virol, 2007. 81 (10): p. 5257-69; In this study, we investigated the effect of CD3 + T cells on the expression of CD3 + T cells. J Immunol, 2007. 179 (7): p. 4741-53].

In addition, the study suggests that the use of common immunogens may increase the extent of the cellular immune response compared to native antigen alone [Yan, J., et al., Enhanced cellular immune responses elicited by an engineered HIV-1 subtype B consensus-based envelope DNA vaccine. Mol Ther, 2007. 15 (2): p. 411-21; Rolland, M., et al., Reconstruction and function of ancestral center-of-tree human immunodeficiency virus type 1 proteins. J Virol, 2007. 81 (16): p. 8507-14]. However, it is recognized that destroying immune tolerance to cancer antigens and making autoimmunity are the main obstacles to cancer vaccination.

There remains a need for compositions useful for inducing an immune response against nucleic acid constructs encoding prostate cancer antigens and prostate cancer antigens and thus destroying immune tolerance. There remains a need for effective preventive and therapeutic vaccines against cost-effective and effective prostate cancer.

An embodiment of the present invention is a composition comprising: a) Amino acid 69, 78, 80, 82, 102, 110, 137, 139, 165, 189, 203, 220, 232 and 248 of SEQ ID NO: 2 are conserved; Or an immunogenic fragment of SEQ ID NO: 2 comprising an amino acid corresponding to at least a 256 amino acid residue of SEQ ID NO: 2, with the proviso that the amino acid 69, 78, 80, 82, 102, 110, 137, 139, 165, 189 , 203, 220, 232, and 248 are preserved; b) SEQ ID NO: 4; A protein that is 98% homologous to SEQ ID NO: 4, provided that the amino acid 21, 86, 127, 129, 154, 156, 182, 195, 206, 218, 220, 237, 249, 255, 265, 271, 275 is preserved; Or an immunogenic fragment of SEQ ID NO: 4 comprising an amino acid corresponding to at least 274 amino acid residues of SEQ ID NO: 4, provided that the amino acid 21, 86, 127, 129, 154, 156, 182, 195, 206, 218 , 220, 237, 249, 255, 265, 271 and 275 are preserved; c) SEQ ID NO: 6; Amino acid 14, 15, 32, 47, 58, 79, 111, 157, 223, 320, 350, 475, 499, 569, 613, 624 of SEQ ID NO: 6, 653, 660, 663, 733 and 734 are preserved; Or an immunogenic fragment of SEQ ID NO: 6 comprising an amino acid corresponding to at least 735 amino acid residues of SEQ ID NO: 6, provided that the amino acid sequence of SEQ ID NO: 6 is 14, 15, 32, 47, 58, 79, 111, 157, 223, 320 , 350, 475, 499, 569, 613, 624, 653, 660, 663, 733 and 734 are preserved; d) SEQ ID NO: 8; A protein which is 98% homologous to SEQ ID NO: 8, provided that the amino acid of SEQ ID NO: 8 is 20,30,31,48,63,74,95,127,173,239,336,366,491,155,564,585, 629, 640, 669, 676, 679, 749 and 750 are conserved; Or an immunogenic fragment of SEQ ID NO: 8 comprising an amino acid corresponding to at least the 751 amino acid residue of SEQ ID NO: 8, provided that the amino acid 20,30,31,48,63,74,95,127,173,239 , 336, 366, 491, 515, 564, 585, 629, 640, 669, 676, 679, 749 and 750 are conserved; e) SEQ ID NO: 10; A protein that is 98% homologous to SEQ ID NO: 10; Or an immunogenic fragment of SEQ ID NO: 10 comprising an amino acid corresponding to at least 333 amino acid residues of SEQ ID NO: 10; f) SEQ ID NO: 12; A protein that is 98% homologous to SEQ ID NO: 12; Or an immunogenic fragment of SEQ ID NO: 12 comprising an amino acid corresponding to at least 349 amino acid residues of SEQ ID NO: 12; g) SEQ ID NO: 14; A protein that is 98% homologous to SEQ ID NO: 14; Or an immunogenic fragment of SEQ ID NO: 14 comprising an amino acid corresponding to at least 129 amino acid residues of SEQ ID NO: 14; Or h) a single peptide linked to amino acids 19-131 of SEQ ID NO: 14; A peptide having a single peptide linked to an amino acid sequence that is 98% homologous to amino acids 19 to 131 of SEQ ID NO: 14; Or a protein having a single peptide linked to an immunogenic fragment of amino acids 19 to 131 of SEQ ID NO: 14, at least 110 amino acid residues of SEQ ID NO: 14, and fragments linked to a signal peptide And a nucleic acid molecule comprising a coding sequence for the coding sequence. In some embodiments, the nucleic acid molecule is selected from encoding the proteins a), b), c) or d).

In another aspect, the invention includes a method of treating an individual diagnosed with prostate cancer, comprising administering to the subject a nucleic acid molecule as described herein.

In another embodiment, there is provided a pharmaceutical composition comprising: a) Amino acid 69, 78, 80, 82, 102, 110, 137, 139, 165, 189, 203, 220, 232 and 248 of SEQ ID NO: 2 are conserved; Or an immunogenic fragment of SEQ ID NO: 2 comprising an amino acid corresponding to at least 261 amino acid residues of SEQ ID NO: 2, provided that the amino acid 69, 78, 80, 82, 102, 110, 137, 139, 165, 189 , 203, 220, 232 and 248 are conserved b) SEQ ID NO: 4; A protein that is 98% homologous to SEQ ID NO: 4, provided that the amino acid 21, 86, 127, 129, 154, 156, 182, 195, 206, 218, 220, 237, 249, 255, 265, 271, 275 is preserved; Or an immunogenic fragment of SEQ ID NO: 4 comprising an amino acid corresponding to at least 274 amino acid residues of SEQ ID NO: 4, provided that the amino acid 21, 86, 127, 129, 154, 156, 182, 195, 206, 218 , 220, 237, 249, 255, 265, 271 and 275 are preserved; c) SEQ ID NO: 6; A protein that is 98% homologous to SEQ ID NO: 6, a protein that is homologous to amino acids 14, 15, 32, 47, 58, 79, 111, 157, 223, 320, 350, 475, 499, 569, 613, 624, 653 , 660, 663, 733 and 734 are preserved; Or an immunogenic fragment of SEQ ID NO: 6 comprising an amino acid corresponding to at least 735 amino acid residues of SEQ ID NO: 6, provided that the amino acid sequence of SEQ ID NO: 6 is 14, 15, 32, 47, 58, 79, 111, 157, 223, 320 , 350, 475, 499, 569, 613, 624, 653, 660, 663, 733 and 734 are preserved; d) SEQ ID NO: 8; A protein which is 98% homologous to SEQ ID NO: 8, provided that the amino acid of SEQ ID NO: 8 is 20,30,31,48,63,74,95,127,173,239,336,366,491,155,564,585, 629, 640, 669, 676, 679, 749 and 750 are conserved; Or an immunogenic fragment of SEQ ID NO: 8 comprising an amino acid corresponding to at least the 751 amino acid residue of SEQ ID NO: 8, provided that the amino acid 20,30,31,48,63,74,95,127,173,239 , 336, 366, 491, 515, 564, 585, 629, 640, 669, 676, 679, 749 and 750 are conserved; e) SEQ ID NO: 10; A protein that is 98% homologous to SEQ ID NO: 10; Or an immunogenic fragment of SEQ ID NO: 10 comprising an amino acid corresponding to at least 333 amino acid residues of SEQ ID NO: 10; f) SEQ ID NO: 12; A protein that is 98% homologous to SEQ ID NO: 12; Or an immunogenic fragment of SEQ ID NO: 12 comprising an amino acid corresponding to at least 349 amino acid residues of SEQ ID NO: 12; g) SEQ ID NO: 14; A protein that is 98% homologous to SEQ ID NO: 14; Or an immunogenic fragment of SEQ ID NO: 14 comprising an amino acid corresponding to at least 129 amino acid residues of SEQ ID NO: 14; Or h) a single peptide linked to amino acids 19-131 of SEQ ID NO: 14; A peptide having a single peptide linked to an amino acid sequence that is 98% homologous to amino acids 19 to 131 of SEQ ID NO: 14; Or a fragment having a single peptide linked to the immunogenic fragment of amino acids 19 to 131 of SEQ ID NO: 14, at least 110 amino acid residues of SEQ ID NO: 14, and fragments linked to the signal peptide. In some embodiments, the protein is selected from the group comprising proteins a), b), c) or d).

Some embodiments of the invention include a method of treating a subject diagnosed with prostate cancer, comprising the step of delivering the protein described herein to the subject.

Another aspect of the invention is a pharmaceutical composition comprising a nucleic acid molecule as provided herein and a pharmaceutically acceptable excipient.

Figure 1 shows the results from in vitro translation performed to confirm the expression of PSA and PSMA antigens;
Figure 2A shows immunogenicity data of cells. Cellular immunogenicity of PSA antigen was determined by interferon-gamma ELISpot;
Fig. 2B shows immunogenicity data of the cells. Fig. Cellular immunogenicity of PSA antigen was determined by interferon-gamma ELISpot;
FIGS. 3A-3C show PSA-specific (left panel), PSMA-specific (middle panel) and whole vaccine-specific (right panel) cytokine producing:% IFN gamma producing CD4 + T cells (FIG. 3A); % IL-2 producing CD4 + T cells (Figure 3B); And characterized by CD4 + T cell response by flow cytometry by plotting% TNFa producing CD4 + T cells (Fig. 3C); Fig.
FIGS. 4A-4C show the results of immunohistochemical staining for PSA-specific (left panel), PSMA-specific (middle panel) and whole vaccine-specific (right panel) cytokine producing:% IFN gamma producing CD8 + T cells (FIG. % IL-2 producing CD8 + T cells (Figure 4B); And CD8 + T cell response characterized by flow cytometry by displaying a graph representing% TNFa producing CD8 + T cells (Figure 4C);
Figures 5A-5B show ELISA data for PSA-specific antibodies one week after final immunization. (Figure 5A) PSA IgG end-point potency. (Figure 5B) Representative IgG titers curves.

The present invention relates to a method for the treatment and / or prophylaxis of prostate cancer, comprising administering to a mammalian subject a prodrug of a prostate specific antigen (PSA), a prostate specific membrane antigen (PSMA) (STEAP) and prostate specific stem cell antigen (PSCA) are provided herein. The term " six-transmembrane epithelial antigen "

Prostate cancer antigens described herein are a consensus sequence derived from a pool of homologous antigens across a number of species, including the species to which the vaccine is targeted. Selectives sorted from antigen sequences to form a common phylogenic tree, such as homapathy (H.sapiens (human)), macaca mullata (M.mulatta (ruffled monkey)), and macaques Will be selected based on the proximity of the species on M. fascicularis (Cynomolgus monkey). Common antigens are not identical to native prostate antigens but have at least 85%, and preferably 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity Share. These listed common cancer antigens can destroy tolerance in the targeted species (or cause autoimmunity) and produce an effective immune response against prostate cancer antigens. Methods for making co-cancer antigen-based DNA are provided herein.

An embodiment of the present invention is a composition comprising: a) Amino acid 69, 78, 80, 82, 102, 110, 137, 139, 165, 189, 203, 220, 232 and 248 of SEQ ID NO: 2 are conserved; Or an immunogenic fragment of SEQ ID NO: 2 comprising an amino acid corresponding to at least a 256 amino acid residue of SEQ ID NO: 2, with the proviso that the amino acid 69, 78, 80, 82, 102, 110, 137, 139, 165, 189 , 203, 220, 232, and 248 are preserved; b) SEQ ID NO: 4; A protein that is 98% homologous to SEQ ID NO: 4, provided that the amino acid 21, 86, 127, 129, 154, 156, 182, 195, 206, 218, 220, 237, 249, 255, 265, 271, 275 is preserved; Or an immunogenic fragment of SEQ ID NO: 4 comprising an amino acid corresponding to at least 274 amino acid residues of SEQ ID NO: 4, provided that the amino acid 21, 86, 127, 129, 154, 156, 182, 195, 206, 218 , 220, 237, 249, 255, 265, 271 and 275 are preserved; c) SEQ ID NO: 6; Amino acid 14, 15, 32, 47, 58, 79, 111, 157, 223, 320, 350, 475, 499, 569, 613, 624 of SEQ ID NO: 6, 653, 660, 663, 733 and 734 are preserved; Or an immunogenic fragment of SEQ ID NO: 6 comprising an amino acid corresponding to at least 735 amino acid residues of SEQ ID NO: 6, provided that the amino acid sequence of SEQ ID NO: 6 is 14, 15, 32, 47, 58, 79, 111, 157, 223, 320 , 350, 475, 499, 569, 613, 624, 653, 660, 663, 733 and 734 are preserved; d) SEQ ID NO: 8; A protein which is 98% homologous to SEQ ID NO: 8, provided that the amino acid of SEQ ID NO: 8 is 20,30,31,48,63,74,95,127,173,239,336,366,491,155,564,585, 629, 640, 669, 676, 679, 749 and 750 are conserved; Or an immunogenic fragment of SEQ ID NO: 8 comprising an amino acid corresponding to at least the 751 amino acid residue of SEQ ID NO: 8, provided that the amino acid 20,30,31,48,63,74,95,127,173,239 , 336, 366, 491, 515, 564, 585, 629, 640, 669, 676, 679, 749 and 750 are conserved; e) SEQ ID NO: 10; A protein that is 98% homologous to SEQ ID NO: 10; Or an immunogenic fragment of SEQ ID NO: 10 comprising an amino acid corresponding to at least 333 amino acid residues of SEQ ID NO: 10; f) SEQ ID NO: 12; A protein that is 98% homologous to SEQ ID NO: 12; Or an immunogenic fragment of SEQ ID NO: 12 comprising an amino acid corresponding to at least 349 amino acid residues of SEQ ID NO: 12; g) SEQ ID NO: 14; A protein that is 98% homologous to SEQ ID NO: 14; Or an immunogenic fragment of SEQ ID NO: 14 comprising an amino acid corresponding to at least 129 amino acid residues of SEQ ID NO: 14; An immunogenic fragment of SEQ ID NO: 14 comprising at least 129 amino acid residues of SEQ ID NO: 14; Or h) a single peptide linked to amino acids 19-131 of SEQ ID NO: 14; A peptide having a single peptide linked to an amino acid sequence that is 98% homologous to amino acids 19 to 131 of SEQ ID NO: 14; Or a protein having a single peptide linked to an immunogenic fragment of amino acids 19 to 131 of SEQ ID NO: 14, at least 110 amino acid residues of SEQ ID NO: 14, and fragments linked to a signal peptide And a nucleic acid molecule comprising a coding sequence for the coding sequence. Two common protein sequences for PSA are disclosed: PSA common antigen sequence 1 (SEQ ID NO: 2) and PSA common antigen sequence 2 (SEQ ID NO: 4). Two common protein sequences for PSMA are disclosed: PSMA common antigen sequence 1 (SEQ ID NO: 6) and PSMA common antigen sequence 2 (SEQ ID NO: 8). Two common protein sequences for STEAP (also referred to herein as STEAP1) are disclosed: STEAP common antigen sequence 1 (SEQ ID NO: 10) and STEAP common antigen sequence 2 (SEQ ID NO: 12). One common protein sequence for PSCA is disclosed: PSCA common antigen sequence (SEQ ID NO: 14). SEQ ID NO: 14 contains an IgE signal peptide. In some embodiments, the PSCA common antigen may comprise amino acids 19-113 of SEQ ID NO: 14 linked to a signal sequence other than the IgE signal of SEQ ID NO: 14. In some embodiments, the nucleic acid molecule is selected from those encoding the proteins a), b), c) or d). In another embodiment, the nucleic acid molecule encodes at least one protein selected from the group consisting of proteins a) or b), and at least one protein selected from the group consisting of proteins c) or d) will be.

The nucleic acid molecule further comprises a nucleotide sequence selected from the group consisting of SEQ ID NO: 2; SEQ ID NO: 4; SEQ ID NO: 6; SEQ ID NO: 8; SEQ ID NO: 10; SEQ ID NO: 12; Or SEQ ID NO: 14; And preferably, SEQ ID NO: 2; SEQ ID NO: 4; SEQ ID NO: 6; Or a molecule encoding at least one protein selected from the group consisting of SEQ ID NO: 8. In some embodiments, the nucleic acid molecule may encode at least one selected from SEQ ID NO: 2 or SEQ ID NO: 4, and at least one protein selected from the group comprising at least one selected from SEQ ID NO: 6 or SEQ ID NO:

In another embodiment, there is provided a pharmaceutical composition comprising: a) Amino acid 69, 78, 80, 82, 102, 110, 137, 139, 165, 189, 203, 220, 232 and 248 of SEQ ID NO: 2 are conserved; Or an immunogenic fragment of SEQ ID NO: 2 comprising an amino acid corresponding to at least a 256 amino acid residue of SEQ ID NO: 2, with the proviso that the amino acid 69, 78, 80, 82, 102, 110, 137, 139, 165, 189 , 203, 220, 232, and 248 are preserved; b) SEQ ID NO: 4; A protein that is 98% homologous to SEQ ID NO: 4, provided that the amino acid 21, 86, 127, 129, 154, 156, 182, 195, 206, 218, 220, 237, 249, 255, 265, 271, 275 is preserved; Or an immunogenic fragment of SEQ ID NO: 4 comprising an amino acid corresponding to at least 274 amino acid residues of SEQ ID NO: 4, provided that the amino acid 21, 86, 127, 129, 154, 156, 182, 195, 206, 218 , 220, 237, 249, 255, 265, 271 and 275 are preserved; c) SEQ ID NO: 6; Amino acid 14, 15, 32, 47, 58, 79, 111, 157, 223, 320, 350, 475, 499, 569, 613, 624 of SEQ ID NO: 6, 653, 660, 663, 733 and 734 are preserved; Or an immunogenic fragment of SEQ ID NO: 6 comprising an amino acid corresponding to at least 735 amino acid residues of SEQ ID NO: 6, provided that the amino acid sequence of SEQ ID NO: 6 is 14, 15, 32, 47, 58, 79, 111, 157, 223, 320 , 350, 475, 499, 569, 613, 624, 653, 660, 663, 733 and 734 are preserved; d) SEQ ID NO: 8; A protein which is 98% homologous to SEQ ID NO: 8, provided that the amino acid of SEQ ID NO: 8 is 20,30,31,48,63,74,95,127,173,239,336,366,491,155,564,585, 629, 640, 669, 676, 679, 749 and 750 are conserved; Or an immunogenic fragment of SEQ ID NO: 8 comprising an amino acid corresponding to at least the 751 amino acid residue of SEQ ID NO: 8, provided that the amino acid 20,30,31,48,63,74,95,127,173,239 , 336, 366, 491, 515, 564, 585, 629, 640, 669, 676, 679, 749 and 750 are conserved; e) SEQ ID NO: 10; A protein that is 98% homologous to SEQ ID NO: 10; Or an immunogenic fragment of SEQ ID NO: 10 comprising an amino acid corresponding to at least 333 amino acid residues of SEQ ID NO: 10; f) SEQ ID NO: 12; A protein that is 98% homologous to SEQ ID NO: 12; Or an immunogenic fragment of SEQ ID NO: 12 comprising an amino acid corresponding to at least 349 amino acid residues of SEQ ID NO: 12; g) SEQ ID NO: 14; An immunogenic fragment of SEQ ID NO: 14 comprising a protein that is 98% homologous to SEQ ID NO: 14 or an amino acid corresponding to at least 129 amino acid residues of SEQ ID NO: 14; Or h) a single peptide linked to amino acids 19-131 of SEQ ID NO: 14; A peptide having a single peptide linked to an amino acid sequence that is 98% homologous to amino acids 19 to 131 of SEQ ID NO: 14; Or a fragment having a single peptide linked to an immunogenic fragment of amino acids 19 to 131 of SEQ ID NO: 14, at least 110 amino acid residues of SEQ ID NO: 14, and fragments linked to a signal peptide. In some embodiments, the protein is selected from the group comprising proteins a), b), c) or d). In another embodiment, the protein is one that encodes at least one protein selected from the group consisting of proteins a) or b) and at least one protein selected from the group consisting of proteins c) or d).

The protein further comprises SEQ ID NO: 2; SEQ ID NO: 4; SEQ ID NO: 6; SEQ ID NO: 8; SEQ ID NO: 10; SEQ ID NO: 12; Or SEQ ID NO: 14; And preferably, SEQ ID NO: 2; SEQ ID NO: 4; SEQ ID NO: 6; Or a protein selected from the group consisting of SEQ ID NO: 8. In some embodiments, the protein may be selected from the group consisting of SEQ ID NO: 2 or at least one selected from SEQ ID NO: 4, and at least one selected from SEQ ID NO: 6 or SEQ ID NO:

Nucleic acid coding sequences are designed to improve and optimize expression. The codons used in these nucleic acid molecules were chosen to produce RNA with reduced secondary structure formation due to intramolecular hybridization. Nucleic acid sequences encoding PSA common antigen sequence 1 (SEQ ID NO: 1) and PSA common antigen sequence 2 (SEQ ID NO: 3) are disclosed. Similarly, the PSMA common antigen sequence 1 (nucleotide 1 to 2250 of SEQ ID NO: 5) and the PSMA common antigen sequence 2 (nucleotide 1-2301 of SEQ ID NO: 7 or SEQ ID NO: 7) as well as the STEAP common antigen sequence 1 No. 9), the STEAP common antigen sequence 2 (SEQ ID NO: 11) and the PSCA common antigen sequence (SEQ ID NO: 13). Also provided is a protein that is 98% homologous to SEQ ID NO: 1 and is up to 98% homologous to PSA common antigen sequence 1 (SEQ ID NO: 2) or SEQ ID NO: 2, preferably amino acids 69, 78 And 98% homologous to SEQ ID NO: 3 and encoding PSA common antigen sequence 2 (SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, or a protein that is homologous to SEQ ID NO: 4 up to 98%, preferably an amino acid 21, 86, 127, 129, 154, 156, 182, 195, 206, 218, 220, 237 , 249, 255, 265, 271 and 275 are provided. Likewise, a protein which is 98% homologous to the nucleotide 2250 of SEQ ID NO: 5 and which is up to 98% homologous to PSMA common antigen sequence 1 (SEQ ID NO: 6) or SEQ ID NO: 6, preferably SEQ ID NO: 6 A nucleic acid sequence encoding amino acids 14, 15, 32, 47, 58, 79, 111, 157, 223, 320, 350, 475, 499, 569, 613, 624, 653, 660, 663, 733 and 734, A homologous 98% homologous to the nucleotide 2301 of SEQ ID NO: 7 and 98% homologous to PSMA common antigen sequence 1 (SEQ ID NO: 8) or SEQ ID NO: 8, preferably an amino acid 20 of SEQ ID NO: 8, As well as nucleic acid sequences coding for the amino acid sequences of SEQ ID NOS: 30, 31, 48, 63, 74, 95, 127, 173, 239, 336, 366, 491, 515, 564, 585, 629, 640, 669, 676, 679, 749 and 750 (SEQ ID NO: 10) or SEQ ID NO: 10, which is 98% homologous to SEQ ID NO: 9 and encodes a protein that is up to 98% homologous to STEAP common antigen sequence 1 A nucleotide that is 98% homologous to SEQ ID NO: 11 and encodes a STEAP common antigen sequence 2 (SEQ ID NO: 12) or a protein that is homologous to SEQ ID NO: 12 up to 98% Homologous and encodes a PSCA common antigen sequence (SEQ ID NO: 14) or a protein that is up to 98% homologous to SEQ ID NO: 14. In some embodiments, the nucleic acid molecule encodes a protein comprising an IgE signal peptide (e.g., SEQ ID NO: 3 encoding SEQ ID NO: 4, nucleotide 1 to 2301 SEQ ID NO: 7 encoding SEQ ID NO: 8, SEQ ID NO: 12 SEQ ID < / RTI > No. 11, which encodes SEQ ID NO: 14, and SEQ ID NO: 13, which encodes SEQ ID NO: 14).

A composition comprising a nucleic acid molecule comprising the coding sequence of a separate nucleic acid molecule provided herein may be useful for inducing an immune response against a prostate protein when administered to an animal. Compositions containing one or more of these nucleic acid sequences may be used as a vaccine or vaccine component to immunize prophylactically or therapeutically against prostate cancer. Likewise, a composition comprising a common protein may be useful for inducing an immune response against prostate protein when administered to an animal. A combination of compositions comprising a nucleic acid molecule comprising a coding sequence of a separate nucleic acid molecule provided herein may be useful for inducing an immune response to a prostate protein and may be administered prophylactically or therapeutically May be used as a vaccine or vaccine component for immunization. Likewise, a composition comprising a common protein may be useful for inducing an immune response against prostate protein when administered to an animal. Compositions containing one or more of these common proteins may be used as a vaccine or vaccine component to immunize prophylactically or therapeutically against prostate cancer.

Vaccines comprising the nucleic acid sequences provided herein are provided. In some embodiments, one or more common prostate antigens selected from the group consisting of a common PSA antigen 1, a common PSA antigen 2, a common PSMA antigen 1, a common PSMA antigen 2, a common STEAP antigen 1, a common STEAP antigen 2, Lt; RTI ID = 0.0 > a < / RTI > A method of inducing an immune response using a nucleic acid sequence encoding one or more prostate antigens comprises the steps of: generating a PSA antigen 1, a common PSA antigen 2, a common PSMA antigen 1, a common PSMA antigen 2, a common STEAP antigen 1, a common STEAP antigen 2, ≪ / RTI >

There is provided a vaccine comprising one or more of a common PSA antigen 1, a common PSA antigen 2, a common PSMA antigen 1, a common PSMA antigen 2, a common STEAP antigen 1, a common STEAP antigen 2 and a common PSCA. Methods of inducing an immune response using one or more of a common PSA antigen 1, a common PSA antigen 2, a common PSMA antigen 1, a common PSMA antigen 2, a common STEAP antigen 1, a common STEAP antigen 2, and a common PSCA are also provided.

Methods of protecting individuals against prostate cancer or methods of treating individuals that have been found to have prostate cancer are provided. The method comprises administering to the subject an effective amount of one or more nucleic acid molecules comprising one or more nucleic acid sequences provided herein. In some methods, the delivery of nucleic acid molecules is made possible by electroporation of tissues that contain the targeted tissue or nucleic acid molecules. The nucleic acid sequence is expressed in the cells of the individual, and the immune response is induced against the prostate protein encoded by the nucleic acid sequence.

One. Justice.

The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting. The singular forms as used in this specification and the appended claims include plural referents unless the context clearly dictates otherwise.

In the present specification, for quotations of numerical ranges, the numbers between each are clearly considered with the same degree of precision. For example, for the range of 6 to 9, the numbers 7 and 8 are considered in addition to 6 and 9, and 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, The numbers 6.8, 6, 9, and 7.0 are clearly considered.

a. Adjuvant ( adjuvant )

&Quot; Adjuvant " as used herein refers to any molecule that is added to the DNA plasmid vaccine described herein to enhance the immunogenicity of the DNA plasmid and the antigen encoded by the encoded nucleic acid sequence described herein below .

b. Antibody

As used herein, an "antibody" refers to an antibody, or fragment, fragment or derivative thereof, of a class IgG, IgM, IgA, IgD or IgE such as Fab, F (ab ') 2, Fd, Diabodies, bispecific antibodies, bifunctional antibodies, and derivatives thereof. The antibody may be an antibody, a polyclonal antibody, an affinity purification antibody, or a mixture thereof, isolated from a serum sample of a mammal that exhibits sufficient binding specificity to the desired epitope or sequence derived therefrom.

c. Password sequence

As used herein, "cryptic sequence" or "encoded nucleic acid" refers to a nucleic acid (RNA or DNA molecule) comprising a nucleotide sequence encoding a protein. The coding sequence may further comprise an initiation and termination signal operably linked to a regulatory element comprising a promoter and a polyadenylation signal capable of directing expression in a cell of the individual or the mammal to which the nucleic acid is to be administered.

d. Complement

As used herein, " complement " or " complementarity " refers to a Watson-Crick (e.g., AT / U and CG) or Hoogsteen base pair between a nucleotide or nucleotide analog of a nucleic acid molecule Which means a nucleic acid that can be used.

e. Common or common sequence

As used herein, " common " or " common sequence " refers to a polypeptide sequence based on the analysis of multiple subtype alignments of specific prostate antigens. Nucleic acid sequences encoding a common polypeptide sequence can be prepared. Vaccines comprising a protein comprising a common sequence and / or nucleic acid molecule encoding such a protein may be used to induce extensive immunity against a particular prostate antigen.

f. Electroporation

The term "electroporation", "electro-penetration" or "electro-mechanical enhancement" (EP), as used interchangeably herein, refers to the use of a transmembrane field pulse to induce a micro- ; Their presence allows the plasmid, oligonucleotide, siRNA, drug, ion and water to pass from one side of the cell membrane to the rest.

g. snippet

&Quot; Fragment " as used herein with reference to a nucleic acid sequence refers to a nucleic acid sequence or a portion thereof, which encodes a polypeptide capable of inducing an immune response in a mammal, wherein the polypeptide cross-reacts with a full-length prostate antigen. The fragment may be a DNA fragment selected from at least one of a common amino acid sequence and various nucleotide sequences encoding a construct comprising such a sequence. The DNA fragment may contain a coding sequence for an immunoglobulin leader such as an IgE or IgG sequence. DNA fragments can encode the following protein fragments.

&Quot; Fragment " for a polypeptide sequence refers to a polypeptide capable of eliciting an immune response in a mammal, which is cross-reacted with a prostate antigen, including, for example, PSA, PSMA, STEAP and PSCA.

The human PSA sequence is about 261 amino acids. The fragment of PSA Common Antigen 1 comprises at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%, and preferably 98% or 99 Wherein the fragment comprises at least one of amino acids 69, 78, 80, 82, 102, 110, 137, 139, 165, 189, 203, 220, 232 and 248. A fragment of PSA common antigen 1 may comprise 255, 256, 257, 258, 259 or 260 amino acids of SEQ ID NO: 2, but preferably may comprise more than 256 amino acids. The fragment of PSA Common Antigen 2 comprises at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%, and preferably 98% or 99% Wherein the fragment comprises at least one of amino acids 21, 86, 127, 129, 154, 156, 182, 195, 206, 218, 220, 237, 249, 255, 265, 271 and 275 . All such fragments of PSA common antigen 2 can also optionally contain amino acids 1-17. In some embodiments, the fragment of PSA Common Antigen 2 may optionally comprise one or more of amino acids 1 to 17 and amino acid 18 to amino acid 278, and the fragment of PSA Common Antigen 2 may also comprise one or more of amino acids 275, 256 , 257, 258, 259 or 260 amino acids, but may preferably comprise more than 274 amino acids.

The human PSMA sequence is about 749 to 750 amino acids. The fragment of PSMA Common Antigen 1 has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%, and preferably 98% or 99% %, With the proviso that the fragments comprise amino acids 14, 15, 32, 47, 58, 79, 111, 157, 223, 320, 350, 475, 499, 569, 613, 624, 653, 660, 663, 733, and 734, respectively. The fragment of PSMA common antigen 1 may comprise amino acids 745, 746, 747, 748 or 749 of SEQ ID NO: 6, but preferably more than 735 amino acids. The fragment of PSMA common antigen 2 is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%, and preferably 98% or 99% %, With the proviso that the fragments comprise amino acids 20, 30, 31, 48, 63, 74, 95, 127, 173, 239, 336, 366, 491, 515, 564, 585, 629, 640, 669, 676, 679, 749, and 750, respectively. All such fragments of PSA Common Antigen 2 can also optionally exclude amino acids 1-16. In some embodiments, the fragment of PSA Common Antigen 2 may optionally comprise one or more of amino acids 1-16, amino acids 17-176, and a fragment of PSMA Common Antigen 2 may also comprise 760, 761 of SEQ ID NO: 8 , 762, 763, 764 or 765 amino acids, but may preferably comprise more than 751 amino acids.

The human STEAP sequence is about 339 amino acids. The common STEAP sequence may comprise an amino acid sequence for an immunoglobulin leader such as IgE or IgG. The common STEAP antigen 2 contains 18 amino acid leader sequences instead of methionine at position 1. The fragment of PSMA Common Antigen 2 has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the leader sequence and amino acids 18-356 of SEQ ID NO: And preferably 98% or 99%. The fragment of PSMA common antigen 1 may comprise amino acids 1 to 350, 1 to 351, 1 to 352, 1 to 353, 1 to 354 or 1 to 355 of SEQ ID NO: 12.

The human PSCA sequence is about 114 amino acids. The common STEAP sequence may contain an amino acid sequence for an immunoglobulin leader such as IgE or IgG. The common PSCA antigen contains 18 amino acid leader sequences instead of methionine at position 1. The fragments of the PSCA common antigens comprise at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% of the leader sequence and the amino acids 18-131 of SEQ ID NO: May include 98% or 99%. The fragment of PSMA common antigen 1 may comprise 1-152, 1-126, 1-127, 1-128, 1-12 or 1-130 amino acids of SEQ ID NO: 14.

h. gene Construct

The term " gene construct " as used herein refers to a DNA or RNA molecule comprising a nucleotide sequence encoding a protein. The coding sequence comprises a start and stop signal operably linked to a regulatory component comprising a promoter and a polyadenylation signal capable of directing intracellular expression of the individual to which the nucleic acid molecule is to be administered. As used herein, the term " expressible form " refers to a genetic construct containing a necessary regulatory element operably linked to a coding sequence that encodes the protein so that the coding sequence is expressed when present in the cell of the individual.

i. Homology

The homology of multiple sequence alignments and phylogenetic trees was generated using ClustalW, a general-purpose multiple sequence alignment program for DNA or proteins.

j. sameness

As used herein in the context of two or more nucleic acid or polypeptide sequences, " identical " or " identity " means that the sequence has the specified percentages of the same residue over the specified region. The percentages can be determined by optimally aligning the two sequences, comparing the two sequences over the specified region, determining the number of positions where the same residue occurs in both sequences, obtaining the number of matched positions, and specifying the number of matched positions Divided by the total number of positions in the region of interest, and multiplying the result by 100 to obtain% sequence identity. Where two sequences are of different lengths or alignment results in more than one staggered end, and the specified region of comparison comprises only a single sequence, the residue of a single sequence is included in the denominator but not the numerator of the calculation. When comparing DNA and RNA, thymidine (T) and uracil (U) may be considered the same. The identity can be manipulated or by using a computer sequence algorithm such as BLAST or BLAST 2.0.

k. Immune response

&Quot; Immune response " as used herein refers to the activation of the immune system of a host, e. G., The immune system of a mammal, responsive to the introduction of an antigen, such as a prostate common antigen. The immune response can be in the form of a cell or a humoral response, or both.

l. Nucleic acid

As used herein, "nucleic acid" or "oligonucleotide" or "polynucleotide" means at least two nucleotides covalently linked. The description of the standard strand also specifies the sequence of the complementary strand. Thus, the nucleic acid also includes the single stranded complementary strand depicted. Multiple variants of a nucleic acid can be used for the same purpose as a given nucleic acid. Thus, the nucleic acid also includes substantially the same nucleic acid and its complement. Single strands provide probes capable of hybridizing the target sequence under stringent hybridization conditions. Thus, the nucleic acid also includes probes that hybridize under stringent hybridization conditions.

The nucleic acid may be single-stranded or double-stranded, or may contain a portion of double-stranded and single-stranded sequences. The nucleic acid can be DNA, a genome and a cDNA, an RNA, or a hybrid, and the nucleic acid is selected from the group consisting of deoxyribo- and ribo-nucleotides and uracil, adenine, thymine, cytosine, guanine, inosine, ≪ / RTI > and combinations thereof. The nucleic acid can be obtained by a chemical synthesis method or a recombinant method.

m. Operably connected

As used herein, " operably linked " means that the expression of a gene is under the control of a promoter spatially linked thereto. The promoter may be located 5 '(upstream) or 3' (downstream) of the gene under its control. The distance between the promoter and the gene may be approximately the same as the distance between the promoter and the gene, which is controlled by the gene derived from the promoter. As is known in the art, this change in distance can be accommodated without loss of promoter function.

n. Promoter

As used herein, " promoter " means a synthetic or naturally-derived molecule capable of conferring, activating or enhancing the expression of a nucleic acid in a cell. Promoters may include one or more specific transcription control sequences to further enhance expression / / or to alter spatial expression and / or spatial expression thereof. The promoter may also include a distal enhancer or repressor component, which may be located as many as several thousand base pairs from the start site of transcription. The promoter may be derived from a source comprising viruses, bacteria, fungi, plants, insects and animals. A promoter may be used to control the expression of a gene component constitutively or differentially in response to an external stimulus such as a cell, tissue, or organ in which expression occurs, or a developmental stage in which expression occurs, or an external stimulus such as physiological stress, pathogen, metal ion or derivative . Typical examples of promoters include bacteriophage T7 promoter, bacteriophage T3 promoter, SP6 promoter, lac operator-promoter, tac promoter, SV40 late promoter, SV40 early promoter, RSV-LTR promoter, CMV IE promoter, SV40 early promoter or SV40 late promoter, and CMV IE promoter.

o. Strict hybridization conditions

As used herein, "stringent hybridization conditions" refer to conditions under which a first nucleic acid sequence (eg, a probe) is hybridized to a second nucleic acid sequence (eg, a target), such as in a complex mixture of nucleic acids. Strict conditions are sequence-dependent and will be different in different environments. The stringent conditions can be chosen to be about 5 to 10 占 폚 lower than the thermal melting point (Tm) for the specific sequence at a given ionic strength pH. The Tm may be the temperature (under defined ionic strength, pH and nucleic acid concentration) at which 50% of the complementary probe to the target is hybridized to the target sequence at equilibrium (if the target sequence is present in excess, 50% Is occupied in an equilibrium state). Stringent conditions are those in which the salt concentration is less than about 1.0 M sodium ion at pH 7.0 to 8.3, such as about 0.01 to 1.0 M sodium ion concentration (or other salt), the temperature is a short probe (e.g., about 10 to 50 nucleotides) And at least about 60 [deg.] C for long probes (e.g., greater than about 50 nucleotides). Strict conditions can also be achieved by addition of an instability agent such as formamide. For selective or specific hybridization, the positive signal may be at least 2 to 10 times background hybridization. Representative stringent hybridization conditions include: 50% formamide, 5 x SSC and 1% SDS, incubation at 42 캜, or incubation at 65 캜, 5 x SSC, 1% SDS, 0.2 x SSC at 65 캜, and Wash with 0.1% SDS.

p. Substantially complementary

As used herein, " substantially complementary " refers to any of the following: 8,9,10,11,12,13,14,15,16,17,18,19,20,21,22,23,24,25,30,35 , 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 180, 270, 360, 450, 540, 630, 720, 810, 900, 990, 1080, 1170 65%, 70%, 90%, 90%, 90%, 90%, 90%, 90%, 90%, 90% Means that hybridization is carried out under stringent hybridization conditions of 75%, 80%, 85%, 90%, 95%, 97%, 98% or 99% identical or both sequences.

q. Substantially the same

As used herein, " substantially identical " means that the first sequence is substantially complementary to the complement of the second sequence, it may be 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 180, 270, 360, 450, A nucleic acid sequence having a first sequence over a nucleotide or amino acid region of greater than or equal to 540, 630, 720, 810, 900, 990, 1080, 1170, 1260, 1350, 1440, 1530, 1620, 1710, 1800, 1890, And the second sequence are at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98% or 99% identical.

r. Subtype or serotype

As used herein, " subtype " or " serotype " is interchangeable and refers to a genetic variant of prostate cancer antigen to prostate cancer antigen, so that one subtype (or variant) It is recognized by the immune system.

p. Mutant

As used herein with respect to nucleic acids, " variants " refer to (i) a portion or fragment of a reference nucleotide sequence; (ii) the complement of the reference nucleotide sequence or a portion thereof; (iii) a nucleic acid substantially identical to a reference nucleic acid or a complement thereof; Or (iv) a nucleic acid that hybridizes under stringent conditions to a reference nucleic acid, its complement, or a sequence substantially identical thereto.

&Quot; Variant " for a peptide or polypeptide is different from the amino acid sequence by insertion, deletion or conservative substitution of an amino acid, but retains at least one biological activity. A variant also refers to a protein having substantially the same amino acid sequence as a reference protein having an amino acid sequence having at least one biological activity. It is recognized in the art that conservative substitutions of amino acids, i. E. Replacing amino acids with different amino acids of similar properties (e. G., Hydrophilicity, degree and distribution of charge region) typically involve side-by-side variations. These ancillary changes can be identified by partially considering the hydropathic index of amino acids as understood in the art. Kyte et al., J. Mol. Biol. 157: 105-132 (1982)). The hydrotherapeutic index of amino acids is based on consideration of its hydrophobicity and charge. It is known in the art that amino acids of similar hydrotherapeutic indices can be substituted and still retain protein function. In one embodiment, an amino acid having a hydropathic index of +/- 2 is substituted. The hydrophilicity of amino acids can also be used to denote substitutions leading to proteins that possess biological functions. Consideration of the hydrophilic nature of amino acids for peptides allows the calculation of the greatest local average hydrophilicity of the peptides, which is a useful measure reported to correlate with antigenicity and immunogenicity. U.S. Patent No. 4,554,101, the disclosure of which is incorporated herein by reference. Substitution of amino acids having similar hydrophilicity values can result in peptides that possess biological activity, such as immunogenicity, as is understood in the art. Substitutions may be made by amino acids having hydrophilicity values within ± 2 of each other. Both the hydrophobicity and hydrophilicity index of amino acids are influenced by specific side chains of amino acids. Consistent with the observations, it is understood that amino acid substitutions compatible with biological action depend on the relative similarity of amino acids, such as those shown by hydrophobicity, hydrophilicity, charge, size and other properties, and in particular on the side chain of the amino acid concerned.

t. vector

As used herein, " vector " means a nucleic acid sequence containing the origin of replication. The vector may be a vector, a bacteriophage, a bacterial artificial chromosome, or a yeast artificial chromosome. The vector may be a DNA or RNA vector. The vector may be a self-replicating extrachromosomal vector, preferably a DNA plasmid.

2. Common prostate antigen

Common antigens are provided herein that are capable of eliciting an immune response in mammals against prostate antigens. Common antigens can include epitopes that make a common antigen particularly effective because immunogens for prostate cancer cells can be induced. The common prostate antigen may comprise a full-length translation product, a variant thereof, a fragment thereof or a combination thereof.

Seven different common prostate antigens have been designed. Two of the common prostate antigens are common PSA antigen 1 (SEQ ID NO: 2) and common PSA antigen 2 (SEQ ID NO: 4). Two of the common prostate antigens are common PSMA antigen 1 (SEQ ID NO: 6) and common PSMA antigen 2 (SEQ ID NO: 8). Two of the common prostate antigens are common STEAP antigen 1 (SEQ ID NO: 10) and common STEAP antigen 2 (SEQ ID NO: 12). One of the common prostate antigens is the common PSCA antigen (SEQ ID NO: 14). The protein may comprise a sequence homologous to the prostate antigen, a fragment of the prostate antigen and a protein having a sequence homologous to the prostate antigen.

The common PSA antigen 1 (SEQ ID NO: 2) is approximately 91% homologous to the human PSA sequence, is approximately 95% homologous to M. fascicuaris PSA, and has about 95% homology to Makacamulata PSA About 96% homology. The common PSA antigen 1 differs from the human PSA sequence at amino acids 69, 78, 80, 82, 102, 110, 137, 139, 165, 189, 203, 220, 232 and 248 of SEQ ID NO:

The common PSA antigen 2 (SEQ ID NO: 4) is about 90-91% homologous to the human PSA sequence and is about 95% homologous to Makacapas ciclass PSA and 95% homologous to Makacar mulatta PSA It is gay. The common PSA antigen 2 contains the leader sequence at its N-terminus. The common PSA antigen 2 is also different from the human PSA sequence at amino acids 21, 86, 127, 129, 154, 156, 182, 195, 206, 218, 220, 237, 249, 255, 265, 271 and 275 of SEQ ID NO: Do.

The common PSMA antigen 1 (SEQ ID NO: 6) is approximately 96% homologous to the human PSMA sequence and 94% homologous to macaqua mullata PSMA. The common PSMA antigen 1 comprises amino acids 14, 15, 32, 47, 58, 79, 111, 157, 223, 320, 350, 475, 499, 569, 613, 624, 653, 660, 663, 734 < / RTI >

The common PSMA antigen 2 (SEQ ID NO: 8) is approximately 96% homologous to human PSA sequence and 94% homologous to macaqua mullata PSA. The common PSMA antigen 2 contains a leader sequence at its N-terminus. The common PSMA antigen 2 also comprises amino acids 20, 30, 31, 48, 63, 74, 95, 127, 173, 239, 336, 366, 491, 515, 564, 585, 629, 640, 669, 676 , 679, 749 and 750, respectively.

The common STEAP antigen 1 (SEQ ID NO: 10) is about 94% homologous to some human STEAP sequences and about 99% homologous to other human STEAP sequences. The common STEAP antigen 1 (SEQ ID NO: 10) is also about 94% homologous to Makacamulata PSMA.

The common STEAP antigen 2 (SEQ ID NO: 12) is about 88% homologous to some human STEAP sequences and about 94% homologous to other human STEAP sequences. The common STEAP antigen 2 (SEQ ID NO: 12) is also about 94% homologous to macaqua mullata PSMA. The common STEAP antigen 2 contains a leader sequence at its N-terminus.

The common PSCA antigen (SEQ ID NO: 14) is about 87% homologous to human PSCA. The common PSCA antigen (SEQ ID NO: 14) differs from human PSCA by the inclusion of leader sequences at its N-terminus.

The protein is selected from the group consisting of PSA common antigen sequence 1 (SEQ ID NO: 2), PSA common antigen sequence 2 (SEQ ID NO: 4), PSMA common antigen sequence 1 (SEQ ID NO: 6), PSMA common antigen sequence 2 1 (SEQ ID NO: 10), STEAP common antigen sequence 2 (SEQ ID NO: 12) or PSCA common antigen sequence (SEQ ID NO: 14).

The protein is selected from the group consisting of PSA common antigen sequence 1 (SEQ ID NO: 2), PSA common antigen sequence 2 (SEQ ID NO: 4), PSMA common antigen sequence 1 (SEQ ID NO: 6), PSMA common antigen sequence 2 1 (SEQ ID NO: 10), STEAP common antigen sequence 2 (SEQ ID NO: 12) or PSCA common antigen sequence (SEQ ID NO: 14).

As noted above, some embodiments include leader sequences at the N-terminus. In some embodiments, the leader sequence is an IgE leader sequence of SEQ ID NO: 16. In some embodiments of the protein sequences provided herein, SEQ ID NO: 16 is removed from it. Likewise, in some embodiments of the protein sequences provided herein, SEQ ID NO: 15 (which encodes SEQ ID NO: 16) is removed therefrom.

Accordingly, some embodiments relate to a protein comprising a signal peptide linked to SEQ ID NO: 2, SEQ ID NO: 6 or SEQ ID NO: 10 instead of the N-terminal methionine presented in the claims (the coding sequence of the signal peptide is typically N Including a start codon that encodes the terminal methionine). Some embodiments relate to a protein comprising a signal peptide linked to amino acids 19-131 of SEQ ID NO: 14. Some embodiments relate to a protein comprising a signal peptide linked to a protein that is 98% homologous to SEQ ID NO: 2, with the proviso that amino acids 69, 78, 80, 82, 102, 110, 137, 139, 165 , 189, 203, 220, 232 and 248 are preserved. Some embodiments relate to a protein comprising a signal peptide linked to a protein that is 98% homologous to SEQ ID NO: 6, with the proviso that amino acids 14, 15, 32, 47, 58, 79, 111, 157, 223 , 320, 350, 475, 499, 569, 613, 624, 653, 660, 663, 733 and 734 are preserved. Some embodiments relate to a protein comprising a signal peptide linked to a protein that is 98% homologous to SEQ ID NO: 10. In each example, when the signal peptide is linked at the N-terminus, it is linked instead of the N-terminus methionine presented in the claims (the coding sequence of the signal peptide typically includes the start codon encoding the N-terminal methionine). Some embodiments relate to a protein comprising a signal peptide linked to a protein that is 98% homologous to amino acids 19-131 of SEQ ID NO: 14. Some embodiments relate to a protein comprising a signal peptide linked to an immunogenic fragment of SEQ ID NO: 2 comprising an amino acid corresponding to at least a 256 amino acid residue of SEQ ID NO: 2, with the proviso that the amino acid 69, 78, 80 , 82, 102, 110, 137, 139, 165, 189, 203, 220, 232 and 248 are preserved. Some embodiments relate to a protein comprising a signal peptide linked to an immunogenic fragment of SEQ ID NO: 6 comprising an amino acid corresponding to at least 735 amino acid residues of SEQ ID NO: 6, with the proviso that amino acids 14, 15, 32 , 47, 58, 79, 111, 157, 223, 320, 350, 475, 499, 569, 613, 624, 653, 660, 663, 733 and 734 are preserved. Some embodiments are directed to a protein comprising a signal peptide linked to an immunogenic fragment of SEQ ID NO: 10 comprising an amino acid corresponding to at least 333 amino acid residues of SEQ ID NO: 10. Some embodiments relate to a protein comprising a signal peptide linked to a protein having a signal peptide linked to an immunogenic fragment of amino acids 19-131 of SEQ ID NO: 14, wherein the fragment comprises at least 110 amino acid residues of SEQ ID NO: 14.

3. Gene sequences, constructs and plasmids

Nucleic acid molecules encoding a common amino acid sequence have been engineered to optimize stability and expression in humans. In particular, codon selection was determined using codons that were based on efforts to minimize intramolecular interactions and secondary structure formation as well as improved expression. The vaccine may comprise one or more nucleic acid sequences that encode one or more of the common forms of immunogenic proteins selected from this group, in order to optimize stability and expression in humans. A nucleic acid sequence is generated that contains the coding sequence for the IgE leader at the 5 'end of the optimized common coding nucleic acid sequence, which encodes a protein having an IgE leader sequence at the N terminus of the common amino acid sequence. In some embodiments, the nucleic acid sequence encoding the IgE leader is SEQ ID NO: 15.

PSA common antigen sequence 1 (protein sequence SEQ ID No. 2; nucleic acid sequence SEQ ID No. 1), PSA common antigen sequence 2 (protein sequence SEQ ID No. 4, nucleic acid sequence SEQ ID No. 3), PSMA common antigen sequence 1 (protein sequence SEQ ID No. 6; (Nucleic acid sequence having nucleotide 1 to 2250 of SEQ ID NO: 5), PSMA common antigen sequence 2 (protein sequence SEQ ID NO: 8, nucleotide sequence having nucleotide 1 to 2301 of SEQ ID NO: 7), STEAP common antigen sequence 1 (SEQ ID NO: 10), STEAP common antigen sequence 2 (protein sequence SEQ ID NO: 12; nucleic acid sequence SEQ ID NO: 11) or PSCA common antigen sequence (protein sequence SEQ ID NO: 14; nucleic acid sequence SEQ ID NO: 13) / RTI > Nucleic Acid Sequence Encoding PSMA Common Antigen Sequence 1 SEQ ID NO: 5 contains, in addition to the PSMA encoding the nucleotide, an additional 9 codons (27) immediately preceding the stop codon encoding the HA tag (SEQ ID NO: 32) Nucleotides). The HA tag is a peptide sequence corresponding to the influenza epitope particularly useful for the detection of proteins. SEQ ID NO: 5 encodes SEQ ID NO: 6 plus an additional nine amino acid sequence SEQ ID NO: 32 linked to its N-terminus for the C-terminus of SEQ ID NO: 6. In some embodiments, the PSMA-1 common antigen is encoded by SEQ ID NO: 5 and comprises a protein having the amino acid sequence of SEQ ID NO: 6 linked at its C-terminus to the N-terminus of SEQ ID NO: In some embodiments, the PSMA-1 common antigen is encoded by nucleotides 1 through 2250 of SEQ ID NO: 5 and comprises a protein having the amino acid sequence of SEQ ID NO: 6. The coding sequence with nucleotides 1-250 of SEQ ID NO: 5 has at least one stop codon at its 3 ' end. Nucleic Acid Sequence Encoding PSMA Common Antigen Sequence 2 SEQ ID NO: 7 contains, in addition to the nucleotides encoding the IgE signal linked to PSMA, an additional (SEQ ID NO: 32) immediately preceding the stop codon encoding the HA tag (SEQ ID NO: 32) Includes 9 codons (27 nucleotides). SEQ ID NO: 7 encodes SEQ ID NO: 8 plus an additional 9 amino acid sequence of SEQ ID NO: 32 linked at its N-terminus to the C-terminus of SEQ ID NO: In some embodiments, the PSMA-2 common antigen is encoded by SEQ ID NO: 7 and comprises a protein having the amino acid sequence of SEQ ID NO: 8 at its C-terminus to the N-terminus of SEQ ID NO: 32. In some embodiments, the PSMA-2 common antigen is encoded by nucleotides 1 to 2301 of SEQ ID NO: 7 and comprises a protein having the amino acid sequence of SEQ ID NO: 8. The coding sequence with nucleotides 1-2301 of SEQ ID NO: 7 has at least one stop codon at its 3'end.

The isolated nucleic acid molecule is selected from the group consisting of PSA common antigen sequence 1 (SEQ ID NO: 2), PSA common antigen sequence 2 (SEQ ID NO: 4), PSMA common antigen sequence 1 (SEQ ID NO: 6), PSMA common antigen sequence 2 It is possible to encode a protein having a sequence which is 98% homologous to the common antigen sequence 1 (SEQ ID NO: 10), STEAP common antigen sequence 2 (SEQ ID NO: 12) or PSCA common antigen sequence (SEQ ID NO: 14) Amino acids 69, 78, 80, 82, 102, 110, 137, 139, 165, 189, 203, 220, 232 and 248 of SEQ ID NO: 2 are conserved and amino acids 21, 86, 127, 129, 154, 156 , 182, 195, 206, 218, 220, 237, 249, 255, 265, 271 and 275 are conserved and amino acids 14, 15, 32, 47, 58, 79, 111, 157, 223, 320 , 350, 475, 499, 569, 613, 624, 653, 660, 663, 733 and 734 are conserved and 20, 30, 31, 48, 63, 74, 95, 127, 173, 239, 336, 366, 491, 515, 564, 585, 629, 640, 669, 676, 679, 749 and 750 are preserved.

The isolated nucleic acid molecule is selected from the group consisting of PSA common antigen sequence 1 (SEQ ID NO: 2), PSA common antigen sequence 2 (SEQ ID NO: 4), PSMA common antigen sequence 1 (SEQ ID NO: 6), PSMA common antigen sequence 2 A protein having a sequence that is 99% homologous to the common antigen sequence 1 (SEQ ID NO: 10), STEAP common antigen sequence 2 (SEQ ID NO: 12) or PSCA common antigen sequence (SEQ ID NO: 14) Amino acids 69, 78, 80, 82, 102, 110, 137, 139, 165, 189, 203, 220, 232 and 248 of SEQ ID NO: 2 are conserved and amino acids 21, 86, 127, 129, 154, 156 , 182, 195, 206, 218, 220, 237, 249, 255, 265, 271 and 275 are conserved and amino acids 14, 15, 32, 47, 58, 79, 111, 157, 223, 320 , 350, 475, 499, 569, 613, 624, 653, 660, 663, 733 and 734 are conserved and amino acids 20, 30, 31, 48, 63, 74, 95, 127, 173, 239 , 336, 366, 491, 515, 564, 585, 629, 640, 669, 676, 679, 749 and 750 are preserved.

The isolated nucleic acid molecules are selected from the group consisting of PSA common antigen sequence 1 (SEQ ID NO: 1), PSA common antigen sequence 2 (SEQ ID NO: 3), PSMA common antigen sequence 1 (SEQ ID NO: 5 or preferably nucleotides 1 to 2250 of SEQ ID NO: PSMA common antigen sequence 2 (SEQ ID No. 7 or preferably nucleotide 1 to 2301 of SEQ ID No. 7), STEAP common antigen sequence 1 (SEQ ID No. 9), STEAP common antigen sequence 2 (SEQ ID No. 11) or PSCA common antigen sequence SEQ ID NO: 13). ≪ / RTI >

The isolated nucleic acid molecules are selected from the group consisting of PSA common antigen sequence 1 (SEQ ID NO: 1), PSA common antigen sequence 2 (SEQ ID NO: 3), PSMA common antigen sequence 1 (SEQ ID NO: 5 or preferably nucleotides 1 to 2250 of SEQ ID NO: PSMA common antigen sequence 2 (SEQ ID No. 7 or preferably nucleotide 1 to 2301 of SEQ ID No. 7), STEAP common antigen sequence 1 (SEQ ID No. 9), STEAP common antigen sequence 2 (SEQ ID No. 11) or PSCA common antigen sequence SEQ ID NO: 13) with a sequence that is 99% homologous.

The isolated nucleic acid molecule can encode a protein comprising the leader sequence at the N-terminus. In some embodiments, the nucleic acid molecule can encode an IgE leader sequence of SEQ ID NO: 16. In some embodiments, the isolated nucleic acid molecule can encode a protein comprising a signal peptide linked to SEQ ID NO: 2, SEQ ID NO: 6 or SEQ ID NO: 10 instead of the N-terminal methionine presented in the claims Sequence typically includes a start codon that encodes an N-terminal methionine). In some embodiments, the isolated nucleic acid molecule can encode a protein comprising a signal peptide linked to amino acids 19-131 of SEQ ID NO: 14. In some embodiments, the isolated nucleic acid molecule can encode a protein comprising a signal peptide linked to a protein that is 98% homologous to SEQ ID NO: 2, with the proviso that amino acids 69, 78, 80, 82, 102 , 110, 137, 139, 165, 189, 203, 220, 232 and 248 are preserved. In some embodiments, the isolated nucleic acid molecule can encode a protein comprising a signal peptide linked to a 98% homologous protein in SEQ ID NO: 6, with the proviso that amino acids 14, 15, 32, 47, 58, 79, 111, 157, 223, 320, 350, 475, 499, 569, 613, 624, 653, 660, 663, 733 and 734 are preserved. In some embodiments, the isolated nucleic acid molecule can encode a protein comprising a signal peptide linked to a protein that is 98% homologous to SEQ ID NO: 10. In an example where a coding sequence for a signal peptide is provided, the signal peptide is linked to a peptide sequence instead of the N-terminal methionine shown in the sequence shown (the coding sequence of the signal peptide typically includes a start codon encoding an N-terminal methionine do). In some embodiments, the isolated nucleic acid molecule can encode a protein comprising a signal peptide linked to a 98% homologous protein to amino acids 19-131 of SEQ ID NO: 14. In some embodiments, the isolated nucleic acid molecule can encode a protein comprising a signal peptide linked to an immunogenic fragment of SEQ ID NO: 2 comprising at least 256 amino acid residues of SEQ ID NO: 2, the corresponding amino acid residue, 2 amino acids 69, 78, 80, 82, 102, 110, 137, 139, 165, 189, 203, 220, 232 and 248 are conserved. In some embodiments, the isolated nucleic acid molecule can encode a protein comprising a signal peptide linked to an immunogenic fragment of SEQ ID NO: 6 comprising an amino acid corresponding to at least 735 amino acid residues of SEQ ID NO: 6, Amino acids 14, 15, 32, 47, 58, 79, 111, 157, 223, 320, 350, 475, 499, 569, 613, 624, 653, 660, 663, 733 and 734 of SEQ ID NO: 6 are conserved. In some embodiments, the isolated nucleic acid molecule can encode a protein comprising a signal peptide linked to an immunogenic fragment of SEQ ID NO: 10, comprising an amino acid corresponding to at least 333 amino acid residues of SEQ ID NO: 10. In some embodiments, the isolated nucleic acid molecule can encode a protein comprising a signal peptide linked to a protein having a signal peptide linked to an immunogenic fragment of amino acids 19 to 131 of SEQ ID NO: 14, At least 110 amino acid residues.

A gene that may comprise a nucleic acid sequence encoding a common prostate antigen described herein that comprises a fragment of a homologous sequence to a common protein sequence, a homologous sequence to a common protein sequence, a common protein sequence, and a fragment of a common protein sequence Constructs are provided herein. Gene constructs can be present in cells as extracellular molecules that work. The gene construct may be a linear mini chromosome comprising a centromere, a telomer or a plasmid or a cosmid.

The genetic construct may also be part of a recombinant viral vector genome including recombinant adenovirus, recombinant adenovirus related virus, and recombinant vaccinia. The genetic construct may be a live attenuated living microbial organism in a cell or part of a genetic material in a recombinant microbial vector.

The genetic construct may comprise a regulatory component for gene expression of the coding sequence of the nucleic acid. The modulating component may be a promoter, an enhancer initiation codon, a stop codon or a polyadenylation signal.

The nucleic acid sequence may constitute a gene construct which may be a vector. The vector can express the antigen in cells of the mammal in an amount effective to induce an immune response in the mammal. The vector may be a recombinant. The vector may comprise a heterologous nucleic acid coding for the antigen. The vector may be a plasmid. The vector may be useful for transfecting a cell with a nucleic acid encoding an antigen, wherein the transfected host cell is cultured and maintained under conditions that result in the expression of the antigen.

In some embodiments, the coding sequence for a single common prostate antigen is provided on a single vector. In some embodiments, the coding sequences for a plurality of common prostate antigens are provided on a single vector. In some embodiments, there is provided a composition comprising a plurality of vectors, a coding sequence for a plurality of common prostate antigens on a plurality of antigens per antigen or vector per vector.

In some embodiments, the coding sequence for two or more different common prostate antigens can be provided on a single vector. In some embodiments, the coding sequence may have distinct promoter control expression. In some embodiments, the coding sequence may have a single promoter control expression by an IRES sequence that separates the coding sequence. The presence of the IRES sequence results in a separate translation of the transcription product. In some embodiments, the coding sequence may have a single promoter control expression by a coding sequence that encodes a proteolytic cleavage peptide sequence that separates the coding sequence of the antigen. A single translation product, which is then treated with a protease that recognizes the protease cleavage site, produces a single translation product to produce a distinct protein molecule. The protease cleavage site used is typically recognized by a protease that is endogenously present in the cell where expression occurs. In some embodiments, a separate coding sequence for the protease may be included to provide for the production of the protease necessary to process the polyprotein translation product. In some embodiments, the vector comprises 1, 2, 3, 4, 5, 6 or all 7 common prostate antigens.

In each and every example presented herein, the coding sequence can be optimized for stability and high levels of expression. In some instances, codons are selected to reduce the secondary structure of RNA, such as those formed due to intramolecular binding.

The vector may comprise a heterologous nucleic acid encoding the antigen and may further comprise an initiation codon, which may be upstream of the antigen coding sequence and the stop codon, and downstream of the antigen coding sequence. The initiation and termination codons may be in frames with the antigen coding sequence. The vector may also comprise a promoter operably linked to an antigen coding sequence. Promoters operably linked to the antigen coding sequence include promoters from simian virus 40 (SV40), mouse mammary tumor virus (MMTV) promoter, human immunodeficiency virus (HIV) Promoter such as bovine immunodeficiency virus (BIV) long terminal repeat (LTR) promoter, Moloney virus promoter, avian leukosis virus (ALV) promoter, cytomegalovirus a cytomegalovirus (CMV) promoter such as a CMV rapid early promoter, an Epstein Barr virus (EBV) promoter or a Rous sarcoma virus (RSV) promoter. The promoter may be a promoter from a human gene such as human actin, human myosin, human hemoglobin, human muscle creatine or human metalothionein. The promoter may also be a natural or synthetic tissue specific promoter, such as a muscle or skin specific promoter. Examples of such promoters are described in U.S. Patent Publication No. 20040175727, the contents of which are incorporated herein by reference in their entirety.

The vector may also comprise a polyadenylation signal, which may be downstream of the common prostate antigen coding sequence. The polyadenylation signal may be an SV40 polyadenylation signal, an LTR polyadenylation signal, a bovine growth hormone (bGH) polyadenylation signal, a human growth hormone (hGH) polyadenylation signal, or a human β- May be a globin polyadenylation signal. The SV40 polyadenylation signal may be a polyadenylation signal from the pCEP4 vector (Invitrogen, San Diego, Calif.).

The vector may also comprise an enhancer upstream of the common prostate antigen coding sequence. Enhancers may be required for DNA expression. The enhancer may be derived from human actin, human myosin, human hemoglobin, human muscle creatine or a virus enhancer such as CMV, HA, RSV or EBV. Polynucleotide functions are described in U.S. Patent Nos. 5,593,972, 5,962,428 and WO94 / 016737, each of which is incorporated herein by reference in its entirety.

The vector may also contain a mammalian replication origin to maintain the vector outside the chromosome and to produce multiple copies of the intracellular vector. The vector may be pVAX1, pCEP4 or pREP4 from Invitrogen (San Diego, Calif.), Which may contain the Epstein-Barr virus origin of replication and the nuclear antigen EBNA-1 coding region and may contain a high copy epitope Cotton replication can be generated. The backbone of the vector may be pAVO242. The vector may be a replication-associated adenovirus type 5 (adenovirus type 5, Ad5) vector.

The vector may also contain regulatory sequences, which may be appropriate for gene expression in mammalian or human cells to which the vector is administered. The common prostate antigen coding sequences may contain codons, which may allow for more efficient transcription of the intracellular coding sequences.

The vector may be pSE420 (Invitrogen, San Diego, Calif.), Which contains Escherichia coli (yikolrayi (E. coli)) can be used to generate the protein. The vector may also be pYES2 (Invitrogen, San Diego, Calif.), Which may be used for protein production in yeast Saccharomyces cerevisiae. The vector may also have the MAXBAC (TM) complete baculovirus expression system (Invitrogen, San Diego, Calif.), Which can be used for protein production in insect cells. The vector can also be pcDNA I or pcDNA3 (Invitrogen, San Diego, Calif.), Which can be used for the production of mammalian cell proteins such as Chinese hamster ovary (CHO) cells. Vectors are generated by readily available starting materials, including routine techniques and procedures, including Sambrook et al., Molecular Cloning and Laboratory Manual, Second Ed., Cold Spring Harbor (1989) Lt; / RTI > vector or system.

The vaccine may comprise one or more of the prostate antigens presented herein and / or the vaccine may comprise one or more nucleic acid sequences encoding one or more of the common prostate antigens selected from this group. The vaccine may comprise one or more of the common prostate antigens presented herein in combination with other immunogenic prostate proteins having sequences other than the common sequences disclosed herein, including natural sequences, and / One or more nucleic acid sequences encoding one or more of the common prostate antigens selected from this group in combination with a nucleic acid molecule encoding another prostate antigen having a sequence other than the sequence.

PSA antigen 1, common, PSA antigen 2, common, PSMA (not shown), prostate cancer cells not restricted by scientific theory, and widely used to induce immune responses (humoral, cellular or both) One or more of the following nucleic acid sequences encoding one or more proteins selected from the group consisting of: antigen 1, common, PSMA antigen 2, common STEAP antigen 1, common STEAP antigen 2, and common PSCA antigen 1. The coding sequence may also include those provided herein including homologous sequences, fragments and homologous sequences of the fragments.

Some embodiments provide a method of generating an immune response against a prostate cancer cell, comprising collectively administering to the subject one or more compositions comprising one or more coding sequences or combinations described herein. Some embodiments provide a method for prophylactically vaccinating a subject against prostate cancer, including, in general, one or more compositions comprising one or more coding sequences or combinations described herein. Some embodiments provide a method of therapeutically vaccinating a subject having prostate cancer, the composition comprising one or more compositions, collectively comprising one or more coding sequences or combinations described herein.

4. Pharmaceutical composition

There is provided herein a pharmaceutical composition according to the invention comprising from about 1 nanogram to about 10 mg of DNA. In some embodiments, the pharmaceutical composition according to the present invention comprises 1) at least 10,15,20,25,30,35,40,45,50,55,60,65,70,75,80,85,90, 95 or 100 nanograms or at least 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 185, 190, 195, 200, 205, 210, 215, 220, 235, 240, 245, 250, 255, 260, 265, 270, 275, 280, 285, 290, 295, 300, 305, 310, 315, 320, 325, 330, 335, 340, 345, 350, 355, 440, 445, 450, 455, 460, 465, 470, 475, 480, 420, 425, 645, 660, 665, 670, 675, 680, 685, 690, 695, 700, 705, 610, 610, 630, 635, 640, 645, 650, 655, 710, 715, 720, 725, 730, 735, 740, 745, 750, 755, 760, 765, 770, 775, 780, 785, 790, 795, 800, 805, 810, 815, 820, 825, 830, 835, 840, 845, 850, 855, 860, 865, 870, 875, 880, 885, 890, 895, 900, 905, 910, 915, 920 , 925, 930, 935, 940, 945, 950, 955, 960, 965, 970, 975, 980, 985, 990, 995 or 1000 micrograms or at least 1.5, , 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5 or 10 mg or more; And 2) up to 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95 or 100 nanograms, , 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, , 150, 155, 160, 165, 170, 175, 180, 185, 190, 195, 200, 205, 210, 215, 220, 225, 230, 235, 240, 245, 250, 255, 260, , 275, 280, 285, 290, 295, 300, 305, 310, 315, 320, 325, 330, 335, 340, 345, 350, 355, 360, 365, 370, 375, 380, 385, 390, 395 , 400, 405, 410, 415, 420, 425, 430, 435, 440, 445, 450, 455, 460, 465, 470, 475, 480, 485, 490, 495, 500, 605, 610, 615, 620 , 625, 630, 635, 640, 645, 650, 655, 660, 665, 670, 675, 680, 685, 690, 695, 700, 705, 710, 715, 720, 725, 730, 735, 740, 745 , 750, 755, 760, 765, 770, 775, 780, 785, 790, 795, 800, 805, 810, 815, 820, 825, 830, 835, 840, 845, 850, 855, 860, , 875, 880, 885, 890, 895, 900, 905, 910, 915, 920, 925, 930, 935, 940, 945, 950, 955, 960, 965, 970, 975, 980, 985, 990, 995 , or 1000 m Or up to 12, or 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5 or 10 mg. In some embodiments, the pharmaceutical composition according to the present invention comprises about 5 nanograms to about 10 milligrams of DNA. In some embodiments, the pharmaceutical composition according to the present invention comprises about 25 nanograms to about 5 milligrams of DNA. In some embodiments, the pharmaceutical composition contains from about 50 nanograms to about 1 milligram of DNA. In some embodiments, the pharmaceutical composition contains about 0.1 to about 500 micrograms of DNA. In some embodiments, the pharmaceutical composition contains about 1 to about 350 micrograms of DNA. In some embodiments, the pharmaceutical composition contains about 5 to about 250 micrograms of DNA. In some embodiments, the pharmaceutical composition contains about 10 to about 200 micrograms of DNA. In some embodiments, the pharmaceutical composition contains about 15 to about 150 micrograms of DNA. In some embodiments, the pharmaceutical composition contains about 20 to about 100 micrograms of DNA. In some embodiments, the pharmaceutical composition contains about 25 to about 75 micrograms of DNA. In some embodiments, the pharmaceutical composition contains about 30 to about 50 micrograms of DNA. In some embodiments, the pharmaceutical composition contains about 35 to about 40 micrograms of DNA. In some embodiments, the pharmaceutical composition contains about 100 to about 200 micrograms of DNA. In some embodiments, the pharmaceutical composition comprises about 10 micrograms to about 100 micrograms of DNA. In some embodiments, the pharmaceutical composition comprises about 20 micrograms to about 80 micrograms of DNA. In some embodiments, the pharmaceutical composition comprises about 25 micrograms to about 60 micrograms of DNA. In some embodiments, the pharmaceutical composition comprises about 30 nanograms to about 50 micrograms of DNA. In some embodiments, the pharmaceutical composition comprises about 35 nanograms to about 45 micrograms of DNA. In some preferred embodiments, the pharmaceutical composition contains about 0.1 to about 500 micrograms of DNA. In some preferred embodiments, the pharmaceutical composition contains about 1 to about 350 micrograms of DNA. In some preferred embodiments, the pharmaceutical composition contains about 25 to about 250 micrograms of DNA. In some preferred embodiments, the pharmaceutical composition contains about 100 to about 200 micrograms of DNA.

The pharmaceutical composition according to the present invention is prepared according to the administration mode to be used. When the pharmaceutical composition is an injectable pharmaceutical composition, they are sterile, non-pyrogenic and non-particulate. An isotonic preparation is preferably used. In general, additives for isotonicity may include sodium chloride, dextrose, mannitol, sorbitol, and lactose. In some cases, isotonic solutions such as phosphate buffered saline are preferred. The stabilizer includes gelatin and albumin. In some embodiments, the vasoconstrictor is added to the formulation.

Preferably, the pharmaceutical composition is a vaccine, more preferably a DNA vaccine.

The vaccine may be a DNA vaccine. DNA vaccines may comprise a plurality of identical or different plasmids comprising a nucleic acid coding sequence for one or more of the common prostate antigens. A DNA vaccine may comprise one or more nucleic acid sequences encoding one or more of the common prostate antigens. When a DNA vaccine comprises more than one common prostate antigen, all such sequences may be present on a single plasmid, or each such sequence may be on a different plasmid.

In some embodiments, the vaccine may comprise a nucleic acid sequence encoding one or more of the common prostate antigens in combination with one or more of the common prostate antigens.

DNA vaccines are disclosed in U.S. Patent Nos. 5,593,972, 5,739,118, 5,817,637, 5,830,876, 5,962,428, 5,981,505, 5,580,859, 5,703,055 and 5,676,594, which are incorporated herein by reference. Are incorporated by reference. The DNA vaccine may further comprise components or reagents that inhibit integration into the chromosome. The vaccine may be an RNA of a prostate antigen. RNA vaccines can be introduced into cells.

The vaccine may be a recombinant vaccine comprising the gene constructs or antigens described above. The vaccine may also comprise one or more common prostate antigens in the form of one or more proteinase subunits or one or more attenuated viral particles comprising one or more common prostate antigens. A vaccine vaccine may be a vaccine, a subunit, and a glycoprotein vaccine as well as an attenuated live vaccine, a death vaccine, and a vaccine that delivers a foreign gene encoding one or more common prostate antigens using a recombinant vector. Examples of attenuated live vaccines include the use of recombinant vectors to deliver prostate antigens, subunit vaccines and glycoprotein vaccines are described in U.S. Patent Nos. 4,510,245; 4,797,368; 4,722,848; 4,790,987; 4,920,209; 5,017,487; 5,077,044; 5,110,587; 5,112,749; 5,174,993; 5,223,424; 5,225,336; 5,240,703; 5,242,829; 5,294,441; 5,294,548; 5,310,668; 5,387,744; 5,389,368; 5,424, 065; 5,451,499; 5,453, 364; 5,462,734; 5,470,734; 5,474, 935; 5,482,713; 5,591, 439; 5,643,579; 5,650,309; 5,698,202; 5,955,088; 6,034, 298; 6,042,836; 6,156,319 and 6,589,529, each of which are incorporated herein by reference.

The provided vaccine may be used to induce an immune response involving a therapeutic or prophylactic immune response. Antibodies and / or killer T cells associated with a common prostate antigen may be produced. Such antibodies and cells can be isolated.

The vaccine may further comprise a pharmaceutically acceptable excipient. Pharmaceutically acceptable excipients can be functional molecules as vehicles, adjuvants, carriers or diluents. The pharmaceutically acceptable excipient may be a facilitating agent, which may include a surfactant, such as an immune-stimulating complexes (ISCOMS), a Freund's incomplete adjuvant, a monophosphoryl lipid A Hyaluronic acid, lipids, liposomes, calcium ions, viral proteins, polyanions, multiple cations or nanoparticles or accelerators, such as liposomes, LPS analogs, muramyl peptides, quinone analogs, squalene and squalene.

Transfection enhancers are multiple cations or lipids, including polyanions, poly-L-glutamate (LGS). The transfusion enhancer is poly-L-glutamate, and more preferably poly-L-glutamate is present in the vaccine at a concentration of less than 6 mg / ml. Transfection enhancers also include vehicles such as surface active agents such as the immunostimulatory complex (ISCOMS), Freund's incomplete adjuvant, LPS analogs including monophosphoryl lipid A, muramyl peptide, quinone analogs, squalene and squalene And hyaluronic acid may also be administered with the gene construct. In some embodiments, the DNA vector vaccine may also comprise a liposome comprising a transfection enhancer such as a lipid, a lecithin liposome as a DNA-liposome mixture (see, for example, W09324640) or other liposomes known in the art, calcium ions, Multiple anions, multiple cations or nanoparticles, or other known transfection enhancers. Preferably, the transfection promoter is a multiple cation or lipid comprising a polyanion, poly-L-glutamate (LGS). The concentration of the transfection agent in the vaccine is less than 4 mg / ml, less than 2 mg / ml, less than 1 mg / ml, less than 0.750 mg / ml, less than 0.500 mg / ml, less than 0.250 mg / Less than 0.050 mg / ml or less than 0.010 mg / ml.

The pharmaceutically acceptable excipient may be an adjuvant. The adjuvant may be another gene that is expressed in an alternative plasmid or is delivered as a protein in combination with the plasmid in a vaccine. Adjuvant has been shown to inhibit the growth of atherosclerotic plaques by a variety of mechanisms including alpha-interferon (IFN- alpha), beta interferon (IFN- beta), gamma interferon, platelet derived growth factor (PDGF), TNF alpha, TNF beta, GM- CSF, epidermal growth factor ), Cutaneous T cell-attracting chemokine (CTACK), epithelial thymus-expressed chemokine (TECK), mucosae-associated epithelial chemokine (MEC), IL -12, IL-15, MHC, CD80, CD86, including IL-15 with a deleted signal sequence, and optionally a signal peptide from IgE. Adjuvant has been shown to inhibit IL-12, IL-15, IL-28, CTACK, TECK, platelet derived growth factor (PDGF), TNF ?, TNF ?, GM- CSF, epidermal growth factor , IL-2, IL-4, IL-5, IL-6, IL-10, IL-12, IL-18 or combinations thereof.

Other genes that may be useful adjuvants include those that encode: MCP-1, MIP-la, MIP-1p, IL-8, RANTES, L-selectin, P-selectin, E-selectin, CD34, GlyCAM 1, MadCAM-1, LFA-1, VLA-1, Mac-1, pl50.95, PECAM, ICAM-1, ICAM-2, ICAM-3, CD2, LFA-3, M- , Fas, TNF receptor, Flt, Apo-1, IL-4, IL-4, IL-18, CD40, CD40L, angiogenic growth factor, fibroblast growth factor, IL- Ap-3, AIR, LARD, NGRF, DR4, DR5, KILLER, TRAIL-R2, TRICK2, DR6, caspase ICE, Fos, c-jun, Sp- 1, Ap-2, p38, p65Rel, MyD88, IRAK, TRAF6, IkB, Inactive NIK, SAPK, SAP-1, JNK, Interferon Reaction Genes, NFkB, Bax, TRAIL, TRAILrec, TRAILrecDRC5, TRAIL- R4, RANK, RANK LIGAND, Ox40, Ox40 LIGAND, NKG2D, MICA, MICB, NKG2A, NKG2B, NKG2C, NKG2E, NKG2F, TAP1, TAP2 and functional fragments thereof.

The vaccine may further comprise a gene vaccine promoter as described in U.S. Application No. 021,579, filed April 1, 1994, which is incorporated herein by reference in its entirety.

5. Delivery Method

Methods for delivering a pharmaceutical preparation, preferably a vaccine, and providing a gene construct and a common prostate antigen, comprising an epitope that produces a specific effective immunogen to which an immune response can be directed against a prostate antigen cell is provided herein do. The method of delivery of the vaccine or vaccine may be provided to induce a therapeutic and / or prophylactic immune response. Vaccines can be delivered to an individual to modulate the activity of the mammalian immune system and enhance the immune response.

Upon delivery of the vaccine to the mammal, and consequently upon delivery of the vaccine within the mammalian cell, the transfected cells will express and secrete the corresponding prostate common protein. These secreted proteins or synthetic antigens will be recognized by the immune system, which will initiate an immune response that may include antibodies made against the antigen and specifically T-cell responses to the antigen. In some instances, the mammal vaccinated with the vaccine discussed herein will have a primed immune system. The vaccine may be delivered to an individual to modulate the immune system activity of the individual, thereby enhancing the immune response.

Vaccines can be delivered in the form of DNA vaccines, and methods of delivery of DNA vaccines are described in U.S. Patent Nos. 4,945,050 and 5,036,006, both of which are hereby incorporated by reference in their entirety.

The vaccine may be administered to a mammal to induce an immune response in the mammal. The mammal may be a human, a non-human primate, a cow, a pig, a sheep, a goat, an antelope, a buffalo, a buffalo, a cow, a deer, a hedgehog, an elephant, a llama, an alpaca, a mouse, a rat or a chicken, Or chicken.

a. Combination therapy

The pharmaceutical composition, preferably a vaccine, may be administered in combination with one or more other prostate proteins or genes. The vaccine may be administered in combination with a protein or gene encoding an Adjuvant, which may be administered in combination with an alpha-interferon (IFN-alpha), beta-interferon (IFN- beta), gamma-interferon, IL-12, IL-2, IL-4, IL-5, IL-6, TNF, TNF ?, GM-CSF, epidermal growth factor (EGF), platelet derived growth factor (PDGF) Selectin, E-selectin, CD34, GlyCAM-1, MadCAM-1, IL-10, IL-12, IL-18, MCP-1, MIP- 1, LFA-1, VLA-1, Mac-1, pl50.95, PECAM, ICAM-1, ICAM-2, ICAM-3, CD2, LFA-3, M-CSF, G- IL-7, nerve growth factor, vascular endothelial growth factor, Fas, TNF receptor, Flt, Apo-1, p55, WSL-1, IL-18 mutant forms, CD40, CD40L, angiogenic growth factor, fibroblast growth factor , Ap-3, AIR, LARD, NGRF, DR4, DR5, KILLER, TRAIL-R2, TRICK2, DR6, caspase ICE, Fos, c-jun, Sp-1, , p38, p65Rel, MyD88, IRAK, TRAF6, IkB, inactive NIK, SAPK, SAP-1, JNK, interferon response genes, NFkB, Bax, TRAIL, TRA May include functional fragments of ILREc, TRAILrecDRC5, TRAIL-R3, TRAIL-R4, RANK, RANK LIGAND, Ox40, Ox40 LIGAND, NKG2D, MICA, MICB, NKG2A, NKG2B, NKG2C, NKG2E, NKG2F, TAP1 or TAP2 have.

b. Route of administration

Vaccines may be administered orally, parenterally, sublingually, transdermally, rectally, transmucosally, topically, through inhalation, orally, into the pleura, intravenously, intraarterially, intraperitoneally, subcutaneously, Intranasal, intraperitoneal, intraperitoneal, intranasal, intraspinal and intraarticular, or a combination thereof. For veterinary uses, the compositions may be administered as a suitably acceptable preparation according to normal veterinary practice. The veterinarian can readily determine the most appropriate dosing regimen and route of administration for a particular animal. The vaccine may be administered by a conventional syringe, a needleless injection device, other physical methods such as a " microprojectile bombardment gone gun " or electroporation (" EP "), " hydrodynamic "

Vectors of the vaccine include DNA injection (also referred to as DNA vaccination) with or without electroporation, liposome mediated, nanoparticle facilitated recombinant vectors, such as recombinant adenovirus, recombinant adenovirus related virus, and recombinant vaccinia Lt; RTI ID = 0.0 > mammalian < / RTI > Prostate antigens can be delivered via DNA injection and in vivo electroporation.

c. Electroporation

Administration of the vaccine via electroporation of the plasmid of the vaccine can be performed using an electroporation apparatus that can be configured to deliver the desired tissue of the mammal with an energy pulse effective to cause reversible pores in the cell membrane, The energy pulse is a constant current similar to the preset current input by the user.

In some embodiments, when electroporation is used, the electroporation device may include an electroporation component and an electrode assembly or handle assembly. The electrical perforation component is one of the various components of the electric perforator, including the controller, current waveform generator, impedance tester, waveform logger, input components, status reporting components, communication ports, memory components, power and power switches. One or more may be included or accepted. Electroporation may be accomplished using in vivo electroporation devices, such as the CELLECTRA (TM) EP system (Inovio Pharmaceuticals, Inc., Blue Bell, Pa.) Or elgen electric perforators, to facilitate transfection of cells with the plasmid (Inovio Pharmaceuticals, Inc., Bluebell, Pa.), An Elgen electroporator.

The electric perforation component can act as one component of the electroporation device and the other component is a separate component (or component) that communicates with the electroporation component. The electrical perforation component may act as one or more components of the ceiling perforation device, which may communicate with another component of the electrical perforation device that is separate from the ceiling perforation component. The components of the electroporation device that are present as part of an electromechanical or mechanical device may not be limited as the components may function as separate components communicating with one another or with one another. The electroporation element can generate a constant current in the desired tissue and deliver an energy pulse including a feedback mechanism. The electrode assembly may include an array of electrodes having a plurality of electrodes in a spatial arrangement, wherein the electrode assembly receives energy pulses from the electroporation unit and transfers them to the desired tissue through the electrodes. At least one of the plurality of electrodes is neutral during delivery of the energy pulse, measures the impedance in the desired tissue, and delivers the impedance to the electroporation component. The feedback mechanism can receive the measured impedance and adjust the energy pulse delivered by the electroporation element to maintain a constant current.

The plurality of electrodes can deliver energy pulses in a decentralized pattern. The plurality of electrodes can deliver energy pulses in a dispersed pattern through the control of the electrodes under the programmed sequence, and the programmed sequence is input by the user to the electroporation component. The programmed sequence may comprise a plurality of pulses delivered in a sequence, each pulse of a plurality of pulses being delivered by at least two active electrodes with one neutral electrode measuring impedance, Subsequent pulses are delivered by one of the at least two active electrodes together with one neutral electrode for measuring the impedance.

The feedback mechanism may be performed by hardware or software. The feedback mechanism can be performed by an analog closed-loop circuit. The feedback can occur every 50 μs, 20 μs, 10 μs, or 1 μs, but is preferably real-time feedback or immediate (ie, substantially immediate as determined by the techniques available for reaction time determination). The neutral electrode measures the impedance in the desired tissue and delivers an impedance to the feedback mechanism. The feedback mechanism responds to the impedance and adjusts the energy pulse to maintain a constant current at a value similar to the preset current. The feedback mechanism can maintain a steady and immediately constant current during the delivery of the energy pulse.

Examples of electroporation devices and electroporation methods that can facilitate DNA vaccine delivery of the present invention include those described in U.S. Patent No. 7,245,963 to Draghia-Akli et al., U.S. Patent Application Publication No. 2005/0052630 to Smith et al. , The contents of which are incorporated herein by reference in their entirety. Other electroporation devices and electroporation methods that may facilitate delivery of DNA vaccines include those provided in co-pending and co-pending U.S. Patent Application No. 11/874072, filed October 17, 2007, U.S. Published Patent Application No. 60 / 852,149, filed October 17, and U.S. Patent Application Serial No. 60 / 978,982, filed October 10, 2007, all of which are hereby incorporated by reference in their entirety for all purposes, .

U. S. Patent No. 7,245, 963 to Draghia-Akli discloses a module electrode system and its use for facilitating the introduction of biomolecules into cells of selected tissues in the body or plant. The module electrode system includes a plurality of needle electrodes; Hypodermic needle; An electrical connector for providing a conductive connection from a programmable constant-current pulse controller to a plurality of needle electrodes; And a power source. The operator can grab a plurality of needle electrodes mounted on the support structure and tightly insert them into a selected tissue in the body or plant. The biomolecule is then transferred through the hypodermic needle into the selected tissue. A programmable constant-current pulse controller is activated, and a constant-current electrical pulse is applied to a plurality of needle electrodes. Applied constant-current electrical pulses facilitate the introduction of biomolecules into cells between multiple electrodes. The entire contents of U.S. Patent 7,245,963 are incorporated herein by reference.

U.S. Patent No. 2005/0052630 to Smith et al. Describes an electroporation apparatus that can be used to effectively facilitate the introduction of biomolecules into cells of a selected tissue in a body or plant. An electroporation apparatus includes an electro-kinetic device ("EKD device") whose operation is embodied by software or firmware. The EKD device generates a series of programmable constant-current pulses between the electrodes in the array based on user control and the input of pulse variables, and stores and acquires current waveform data. The electroporation apparatus also includes a replaceable electrode disc having an array of needle electrodes, a central scan channel for the needles, and a removable guide disc. The entire disclosure of U.S. Patent Publication No. 2005/0052630 is incorporated herein by reference.

The electrode arrays and methods described in U.S. Pat. No. 7,245,963 and U.S. Patent Publication No. 2005/0052630 may be suitable for deep penetration into tissues such as muscles as well as other tissues or organs. Because of the configuration of the electrode array, in the section described above by the electrode, the injection needle (to deliver the biomolecule of choice) is also completely inserted into the target organs, and the injection is administered perpendicular to the target tissue. The electrodes described in U.S. Patent No. 7,245,963 and U.S. Patent Publication No. 2005/005263 are preferably 20 mm long and 21 gauge.

In addition, in some embodiments, including an electric piercing device and its use, an electric piercing device is contemplated that is described in the following patents: U.S. Patent 5,273,525 issued December 28, 1993, August 29, 2000 U.S. Patent No. 6,110,161 issued July 17, 2001, U.S. Patent No. 6,261,281 issued October 17, 2001, U.S. Patent No. 6,958,060 issued October 25, 2005, and U.S. Patent No. 6,939,862 issued September 6, 2005. Further, a patent covering such content is provided in U.S. Patent No. 6,697,669, issued on February 24, 2004, which is related to the delivery of DNA using any of a variety of devices, U.S. Patent No. 7,328,064, which describes a method of injecting DNA, is contemplated herein. The above patents are incorporated by reference in their entirety. Another embodiment of an electroporation device for use with the cancer antigen described herein is an Elgen EP device (Inovio Pharmaceuticals, Inc. of Bluebell, Pa.).

d. Vaccine manufacturing method

Methods for the preparation of DNA plasmids comprising the DNA vaccines discussed herein are provided herein. After the final subcloning step into the mammalian expression plasmid, DNA plasmids can be inoculated with cell cultures in large-scale expression tanks using methods known in the art.

The DNA plasmids for use with the EP apparatus of the present invention may be formulated or prepared using a combination of known devices and techniques, but preferably they are prepared as described in an approved, co-pending, USA is prepared using the optimized plasmid production technique described in patent application 60 / 939,792. In some instances, the DNA plasmids used in these studies can be formulated at a concentration of at least 10 mg / ml. The manufacturing technique may also include various devices and protocols commonly known to those skilled in the art, in addition to those described in U.S. Patent No. 60 / 939,792, including those described in U.S. Patent 7,238,522, issued July 3, 2007, Or accept. The above-mentioned applications and patents US 60 / 939,792 and US 7,238,522 are each hereby incorporated by reference in their entirety.

Example

The invention is further illustrated in the following examples. It is to be understood that these embodiments are representative of preferred embodiments of the invention, but are provided by way of example only. From the above discussion and these examples, one skilled in the art can ascertain the essential characteristics of the invention, and various changes and modifications of the invention may be made to various phrases and conditions without departing from the spirit and scope thereof. Accordingly, various modifications of the invention in addition to those shown and described herein will become apparent to those skilled in the art from the foregoing description. Such modifications are also intended to fall within the scope of the appended claims.

Example  One

Common immunogens for PSA and PSMA are described in Laddy, D. J., Yan, J., Corbitt, N., Kobasa, D., Kobinger, G. P., Weiner, (2007). Immunogenicity of the novel consensus-based DNA vaccine against avian influenza. Vaccine. D., Kobinger, GP, Khan, AS, Greenhouse, J., Sardesai, NY, Draghia-Akli, R., and Laddy, DJ, Yan, J., Kutzler, M., Weiner, DB (2008). Heterosubtypic Protection against Pathogenic Human and Avian Influenza Viruses via In Vivo Electroporation of Synthetic Consensus DNA Antigens. PLoS ONE. 3, < / RTI > e2517]. ≪ RTI ID = 0.0 >

The common antigen sequences were synthesized by GeneScript (Fitz-Cathay Way, NJ). The HA tag was included at the C-terminus of the antigen sequence. Antigen sequences were optimized for mRNA stability and codon usage in humans. The final sequence was cloned into the BamHI and XhoI sites of the pVAX1 vector (Invitrogen, Carlsbad, Calif.).

A common PSA antigen 1 (SEQ ID NO: 2) was made. This sequence containing 261 amino acids was compared with each of the PSA sequences presented in Table 1. The PSA sequence used includes two human sequences, a sequence derived from Makakapas cicolaris and a sequence derived from macaques mullata. Table 1 includes sequence numbers and registration numbers for each sequence used in comparison to the common PSA antigen 1 (SEQ ID NO: 2).

SEQ ID NO: Species and proteins Registration Number Number of amino acids For SEQ ID NO: 2 Prize Same sex% 17 Homo sapiens PSA iso1 NP001639.1 261 91 18 Homo sapiens PSA gbAAA60193.1 262 91 19 Do Kaka Paz City Kula lease KLK3 Q6DT45.1 261 95 20 Makaka Mulatta PSA NP001036241.1 p 261 96

Multiple sequence alignments of homo sapiens (SEQ ID NO: 17 and SEQ ID NO: 18), macaqua mulatta (SEQ ID NO: 20) and macacapas ciculacea (SEQ ID NO: 19) PSA sequences were linked to a common PSA antigen 1 . KLK3 (Calicrain 3) is a gene encoding PSA and is a pseudonym with PSA. PSA antigen 1 is 91% homologous to homo sapiens, 96% homologous to macaqua mullata, and 95% homologous to the macaquafas ciclaris whole-length PSA protein sequence.

Example 2

A common PSA antigen 2 (SEQ ID NO: 4) was made. This sequence, containing 279 amino acids, including the IgE leader sequence, was compared with each of the PSA sequences shown in Table 2. The PSA sequence used includes two human sequences, a sequence derived from Makakapas cicolaris and a sequence derived from macaques mullata. Table 2 includes sequence numbers and registration numbers for each sequence used in comparison to the common PSA antigen 2 (SEQ ID NO: 4).

SEQ ID NO: Species and proteins Registration Number Number of amino acids An image for SEQ ID NO: 4 Gay % 17 Homo sapiens PSA iso1 NP001639.1 261 91 18 Homo sapiens PSA gbAAA60193.1 262 90 19 Makaka Paz City Kula lease KLK3 Q6DT45.1 261 95 21 Makakamulata PSA AAZ82258.1 244 95

Multiple sequence alignments of homo sapiens (SEQ ID NO: 17 and SEQ ID NO: 18), macaqua mulatta (SEQ ID NO: 21) and macaquat pacificulosus (SEQ ID NO: 19) . KLK3 (Calicrain 3) is a gene encoding PSA and is a pseudonym with PSA. PSA antigen 1 is 90-91% homologous to homo sapiens, 95% homologous to the Makacapas ciclulatus full-length PSA protein sequence, 95% homologous to the Makacapass ciculous partial PSA protein sequence to be.

Example  3

A common PSMA antigen 1 (SEQ ID NO: 6) was made. This sequence, containing 750 amino acids, was compared with each of the PSMA sequences presented in Table 3. The PSMA sequence used contains two human sequences, a sequence derived from macaques mullata. Table 3 includes sequence numbers and registration numbers for each sequence used in comparison to the common PSMA antigen 1 (SEQ ID NO: 6).

SEQ ID NO: Species and proteins Registration Number Number of amino acids Homology to SEQ ID NO: 6 castle % 22 Homo sapiens PSMA GCPII_iso1 NP_004467.1 750 96 23 Homo sapiens PSMA AAC83972.1 749 96 24 Makaka Other times, mullahs GCPII iso1 XP_001096141.2 735 94

Multiple sequence alignments of Homo sapiens, Makacamulata PSMA sequences were made by common PSMA antigen 1. The PSMA antigen 1 consensus sequence (SEQ ID NO: 6) was 96% homologous to Homo sapiens PSMA (SEQ ID NO: 22 and SEQ ID NO: 23) and 94% homologous to the Makaka mullata battlefield PSMA protein sequence (SEQ ID NO: to be.

Example  4

A common PSMA antigen 2 (SEQ ID NO: 8) was made. This sequence containing 766 amino acids, including the IgE leader sequence, was compared to each of the PSMA sequences presented in Table 4. The PSMA sequence used contains two human sequences, a sequence derived from macaques mullata. Table 4 contains sequence numbers and registration numbers for each sequence used in comparison to the common PSMA antigen 2 (SEQ ID NO: 8).

SEQ ID NO: Species and proteins Registration Number Number of amino acids Homology to SEQ ID NO: 8 castle % 22 Homo sapiens PSMA GCPII_iso1 NP_004467.1 750 96 23 Homo sapiens PSMA AAC83972.1 749 96 24 Makaka Mulatta GCPII isol XP_001096141.2 735 94 25 Makaka Other times, mullahs GCPII iso2 XP_002799784.1 704 94

Multiple sequence alignments of Homo sapiens (SEQ ID NO: 22 and SEQ ID NO: 23), Makacamulata PSMA sequences (SEQ ID NO: 24 and SEQ ID NO: 25) were generated by PSMA antigen 2. The PSMA antigen 2 consensus sequence (SEQ ID NO: 8) is 96% homologous to the Homo sapiens PSMA protein sequence and 94% homologous to the Makacar mulatta PSMA protein sequence.

Example  5

A common STEAP antigen 1 (SEQ ID NO: 10) was made. This sequence, containing 339 amino acids, was compared with each of the STEAP sequences presented in Table 5. The used STEAP sequence contains two full-length human sequences, a full-length sequence from macaques mullata, and two shorter human sequences. Table 5 includes sequence numbers and registration numbers for each sequence used in comparison to common STEAP antigen 1 (SEQ ID NO: 10).

SEQ ID NO: Species and proteins Registration Number Number of amino acids Homology to SEQ ID NO: 10 castle % 26 Homo sapiens STEAP1 NP_036581.1 339 99 27 Homo sapiens STEAP1 Gb_EAL24167.1 339 99 28 Makaka Other times, mullahs STEAP1 XP_001103605.1 339 98 29 Homo sapiens STEAP1 CRA b EAW93751.1 259 94 30 Homo sapiens STEAP1 isoform EAW93749.1 258 94

Multiple sequence alignments of homo sapiens and macaqua mullata STEAP sequences were made by STEAP antigen 1. The STEAP antigen 1 consensus sequence (SEQ ID NO: 10) is 99% homologous to human full length isoforms (SEQ ID NO: 26 and SEQ ID NO: 27), shorter homo sapiens isoform (SEQ ID NO: 29 and SEQ ID NO: 30) And 94% homology to the MACA mullata battlefield STEAP1 protein sequence (SEQ ID NO: 28).

Example  6

A common STEAP antigen 2 (SEQ ID NO: 12) was made. This sequence, containing 356 amino acids, was compared with each of the STEAP sequences presented in Table 6. The used STEAP sequence contains two full-length human sequences, a full-length sequence from macaques mullata, and two shorter human sequences. Table 6 includes sequence numbers and registration numbers for each sequence used in comparison to common STEAP antigen 2 (SEQ ID NO: 12).

SEQ ID NO: Species and proteins Registration Number Number of amino acids Homology to SEQ ID NO: 12% 26 Homo sapiens STEAP1 NP_036581.1 339 94 27 Homo sapiens STEAP1 Gb_EAL24167.1 339 94 28 Makaka Other times, mullahs STEAP1 XP_001103605.1 339 94 29 Homo sapiens STEAP1 CRA b EAW93751.1 259 88 30 Homo sapiens STEAP1 isoform EAW93749.1 258 88

Multiple sequence alignments of homo sapiens and macaqua mullata STEAPl sequences were made by common STEAP1 antigen 2. The STEAP1 antigen 2 consensus sequence (SEQ ID NO: 12) is 94% homologous to human full-length isoforms (SEQ ID NO: 26 and SEQ ID NO: 27), 88% homologous to the shorter homo sapiens isoform (SEQ ID NO: 29 and SEQ ID NO: Homology and 94% homology to the macaqua mullata battle STEAP1 protein sequence (SEQ ID NO: 28).

Example  7

A common PSCA antigen (SEQ ID NO: 14) was made. This sequence, containing 131 amino acids and containing the IgE leader sequence, was compared to the PSCA sequence presented in Table 7. The PSCA sequence used was a full-length human sequence. Table 7 includes sequence numbers and registration numbers for the sequences used in comparison with the common PSCA antigen (SEQ ID NO: 14).

SEQ ID NO: Species and proteins Registration Number Number of amino acids Homology to SEQ ID NO: 14% 31 Homo sapiens PSCA NP_005663.2 114 87

Multiple sequence alignments of the Homo sapiens PSCA sequence (SEQ ID NO: 31) were made by the common PSCA antigen (SEQ ID NO: 14). The PSCA antigen common sequence is 87% homologous to the full length Homo sapiens PSCA.

Example  8

In-vitro translation was performed to confirm the expression of PSA and PSMA antigens. A TNT (registered trademark) attribute coupled transcription / translation system (Quick Coupled Transcription / Translation System) and 35S-methionine (Promega) were used. pVAX vector alone (negative control) or pVAX backbone with PSA or PSMA antigen insert and 35S-methionine was added to the reaction mixture according to the manufacturer's instructions. The reaction was carried out at 30 DEG C for 2 hours. Labeled proteins were immunoprecipitated with anti-HA affinity gel (Sigma, St. Louis, MO, USA) by stirring overnight at 4 째 C in radioimmunoprecipitation assay (RIPA) buffer. Immunoprecipitated proteins were then electrophoresed on fixed and dried SDS-PAGE gels. Expression of the 35S-labeled protein was detected by autoradiography. The results are shown in Fig.

Example  9

The cellular immunogenicity of PSA and PSMA antigens was determined by interferon-gamma ELISpot.

Female, 4-6-week-old BALB / c mice were purchased from Jackson Laboratories (Bar Harbor, Maine). All animals were housed in temperature-controlled, light-duty facilities at the University of Pennsylvania. Animal care was conducted according to the guidelines of the National Institutes of Health and the University of Pennsylvania Institutional Care and Use Committee.

For cell immunogenicity studies, 10 or 20 [mu] g of each antigen was injected intramuscularly followed by electroporation using a CELLECTRA (TM) adaptive constant current device (Inovio Pharmaceuticals, Inc., Blue Bell, PA) / c mice to the tibialis anterior. Mice (n = 5 per group) received immunizations twice in week 0 and week 2. Two 0.1 Amp constant-current square-wave pulses were delivered through a triangular three-electrode array of 26-gauge solid stainless steel electrodes. Each pulse was 52 milliseconds in length by a one second delay between pulses. The mice received a total of 2 immunizations administered 2 weeks apart. Mice were humanely sacrificed 1 week after the second immunization for analysis of cellular and humoral immune responses.

Cells and responses were evaluated one week after final immunization (week 5). ELISpot analysis was used to determine the antigen-specific secretion of IFNy. Bottomed-out Immobilon-P plates (Millipore, Billerica, Mass.) Overnight at 4 ° C using a mouse IFN gamma capture antibody (R & D Systems, Minneapolis, Minn.). Splenocytes were isolated aseptically and cultured in R10 medium (Rosewell Park Memorial Institute medium 1640 supplemented with 10% fetal bovine serum, 1% antibiotic-antifungal and 0.1% 2-mercaptoethanol) Lt; / RTI > 2 x 10 < ⁵ > spleen cells from immunized mice were added to each well of a 96-well plate and either R10 (negative control), Concanavalin A (positive control) (Sigma, St. Louis, MO) Stimulated overnight at 37 < 0 > C, 5% CO2 in the presence of a specific peptide pool. The following day, mouse IFN gamma detection antibody (R & D Systems, Minneapolis, MN) was added to plates incubated overnight at 4 < 0 > C. The following day, streptavidin-ALP (MabTech, Sweden) was added to the plate for 2 hours and the antigen-specific spot visualized with a BCIP / NPT substrate (MabTech, Sweden). PSA and PSMA peptides were 15-mer peptides that extended the entire length of the common immunogen and did not contain the HA tag or leader sequence and were as much as 11 amino acids and were synthesized by GenScript (Fitz-Kataway, NJ) . PSA and PSMA peptides were used at a final concentration of 1.0 [mu] g / ml for each peptide. An antigen-specific cellular response was evaluated one week after the last immunization using an IFN gamma ELISpot. For PSA, the IFN gamma response was similar for 10 μg (772.2 +/- 138.2 SFU) and 20 μg (771.1 +/- 155.2 SFU) vaccine doses (FIG. 2A). In contrast, there was a dose-dependent increase in PSMA-specific FN gamma response by 20 microgram vaccine (1585.0 +/- 194.0 SFU) compared to 10 μg vaccine (1047.2 +/- 160.7 SFU) (Figure 2B) . The minimal background for PSA or PSMA response was observed in naïve mice.

Example  10

IFN gamma, IL -2 and TNF The vaccine-induced CD4 + And CD8 + T cell generation

The immunogenicity of the cells was further characterized by flow cytometry for the joint delivery of PSA and PSMA vaccines. Antigen-specific CD4 + and CD8 + T cell generation of IFNγ, IL-2 and TNFα was determined for the PSA and PSMA components of the total vaccine-specific and total vaccine-specific responses (n = 5).

Cellular immune responses were also determined by flow cytometric staining using intracellular cytokine staining and manufacturer's instructions using a CytoFix / CytoPerm kit (BD Biosciences, San Diego, Calif.). Splenocytes harvested from immunized mice were washed with PBS and resuspended in R10 medium at a final concentration of 107 cells / ml. Cells were seeded into a 96-well round bottom flask in a volume of 100 쨉 l and cultured in a medium containing an additional 100 의 of R10 medium (negative control), antigen-specific peptide pool or phorbol myristate acetate (PMA, 10 ng / ml) and inomycin (250 ng / ml; positive control) (Sigma, St. Louis, MO) were added and the plates were incubated at 37 ° C, 5% CO2 for 6 hours. All stimulation media contained 1 μg / μl of each of GolgiPlug and GolgiStop (BD Biosciences, San Diego, Calif.). Plates were spun down at the end of the incubation period and washed twice with PBS. The cells were then stained with a purple dye for a viability (LIVE / DEAD Violet Viability Dye, Invitrogen, Carlsbad, Calif.) For 30 min at 4 ° C. After washing with PBS as above, the cells were stained externally for 30 minutes with anti-CD4 PerCPCy5.5 and anti-CD8 APC at 4 < 0 > C, then fixed and settled. Anti-IFN? AlexaFluor-700 and anti-TNF? FITC (BD Biosciences, San Diego, Calif.) Were added and incubated again at 4 ° C for 30 minutes. Cells were given a final wash with PBS and fixed in 1% PFA.

Joint delivery of PSA and PSMA vaccines induced strong CD4 + secretion of IFNγ, IL-2 and TNFα. The percentage of PSA-specific (0.21%) and PSMA-specific (0.24%) IFNγ producing CD4 + T cells contributed equally to the overall vaccine-specific CD4 + T cell IFNγ response (0.44%) (FIG. 3A). PSMA-specific CD4 + T cells producing IL-2 (1.08%) contained the majority of the total percentage of CD4 + T cells producing vaccine-specific IL-2 (1.40%) (Fig. 3B). The percentage of CD4 + T cell production of PSA (0.31%) and PSMA (0.29%) induced TNFa contributed equally to the overall vaccine-specific response (0.60%) (FIG. 3C). Overall, the CD4 + T cell response was well balanced between PSA and PSMA, except for PSMA, which induced the majority of vaccine-specific CD4 + T cell IL-2 production.

The vaccine induced strong antigen-specific CD8 + T cell production of IFNy and IL-2 and less TNFa. Both PSA (0.70%) and PSMA (0.67%) induced strong CD8 + T cell IFNγ production. In fact, vaccine-specific CD8 + T cell secretion of IFN gamma contained 1.37% of the total CD8 + T cell population (Fig. 4A). The vaccine also induced a strong CD8 + T cell IL-2 response (1.54%). Similar to the CD4 + T cell IL-2 response, the percentage of PSMA-specific (1.06%) CD8 + T cells secreting IL-2 was approximately 2-fold greater than PSA-specific (0.47%) (FIG. 4B). The overall percentage of vaccine-specific CD8 + T cell generation of TNFa (0.11%) responds to the PSA component of the vaccine (Figure 4C). In summary, there was a high percentage of IFN gamma and IL-2 vaccine-specific CD8 + T cell generation. Similar to the CD4 + T cell response, IFN gamma production was equally balanced between PSA and PSMA, and the magnitude of the IL-2 PSMA-specific response was greater than the magnitude of the PSA-specific response.

Example  11

PSA-specific IgG seroconversion

Antibody response can play an important role in tumor immunotherapy. Thus, the present inventors next tested these parameters of the immune response to PSA antigens based on protein target availability.

To determine the PSA-specific serum antibody titers, 96-well Nunc-Immuno MaxiSorp plates (Nunc, Rochester, NY) were incubated with recombinant PSA protein (Fitzgerald Industries, Inc., Acton, Mass.) Diluted in PBS overnight at 4 ° C 1 占 퐂 / well. Plates were washed with PBS, 0.05% Tween 20 (PBST), blocked with 10% BSA / PBST for 1 hour at room temperature, step diluted with serum from naïve animals immunized for 1 hour at room temperature ≪ / RTI > The plates were then washed three times with PBST and goat anti-mouse IgG (Santa Cruz, Santa Cruz, Calif.) Was added at a dilution of 1: 5,000 in PBST. The bound enzyme was detected by SigmaFAST O-phenylenediamine dihydrochloride (OPD; Sigma-Aldrich, St. Louis, MO) and the optical density was measured using a Biotek (Wincusi, Vt. ≪ / RTI > plate reader at 450 nm. The end-point titre was determined as previously described (Frey, A. et al., 1998). Briefly, the upper prediction limit was calculated using Student's t-distribution. The equation for defining the upper prediction limit is expressed by multiplying the standard deviation by a factor based on the number of negative controls (n = 5) and the confidence level (95%). The end point reversal reported as the reciprocal of the final dilution above the upper predicted limit.

In addition to conferring strong cell-mediated immunity, the PSA vaccine also induced a strong antigen-specific humoral response. Antibody titers were determined by ELISA in serum isolated from mice one week after final immunization (n = 5). The vaccine induced an average PSA-specific antibody end-point titre of 4,427 (ranging from 1581 to 15,811) (Fig. 5A). The persistence of these reactions can be equally important.

Example 12

Prostate specific antigen amino acid sequences available from GenBank include: gb_EAW71923.1_Homo sapiens_klk3_CRAb;

_001639.1_ Homo sapiens _ PSA_iso1_ preproprotein;

gb_AAA59995.1_ Homo sapiens _ PSA_ precursor; gb_AAA60193.1_ Homo sapiens _PSA;

gb_EAW71933.1_ Homo sapiens _klk3_CRA_l;

NP_001025218.1_ Homo sapiens _ PSA_iso3_ preproprotein; gb_CAD54617.1_ Homo sapiens _PSA;

gb_CAD30844.1_ Homo sapiens_PSA; gb_AAA59996.1_ Homo sapiens_PSA_ precursor;

gb_AAD14185.1_ Homo sapiens _PSA; Q6DT45.1_ Makakapasu Cyclicals _KLK3;

NP_001036241.1_ MAKAKA MULLATA _ PSA_ precursor; AAZ82258.1_Macakamula_PSA;

AAZ82255.1_G.gorilla_PSA; gi | 163838666 | ref | NP_001106216.1 | Plasma calicaine [Olive baboons (Papio anubis)]; gi | 73746696 | gb | AAZ82261.1 | Prostate specific antigen [Olive baboons];

i | 73746692 | gb | AAZ82259.1 | Prostate-specific antigen (Erythrocebus patas);

gi | 73746694 | gb | AAZ82260.1 | Prostate-specific antigen [Cercopithecus cephus];

gi | 73746682 | gb | AAZ82254.1 | Prostate-specific antigen (Panopus monkey);

gi | 73746680 | gb | AAZ82253.1 | Prostate-specific antigens (Pan troglodytes);

gi | 73746686 | gb | AAZ82256.1 | Prostate-specific antigen [orangutan (Pongo pygmaeus)]; And

3746688 | gb | AAZ82257.1 | Prostate specific antigen [Nomascus gabriellae].

PSMA amino acid sequences available from GenBank include: NP_004467.1_human_GCPII_iso1; Human _PSMA_AAC83972.1; Makaka mullata_GCPII_iso1 XP_001096141.2; And Makaka Mullata_GCPII_iso2_XP_002799784.1.

The STEAP amino acid sequences available from GenBank include the following: NP036581.1_human _STEAP1; EAL24167.1_Human_STEAP1; XP001103605.1_ MAKACA MULTITA_STEAP1_iso3; EAW93751.1_Human_STEAP1_CRAb; EAW93749.1_Human_STEAP1_CRAa; XP001164838.1_ Chimpanzee (P.troglodytes) _STEAPiso2; XP002818311.1_ Sumatra Orangutan (P.abelii) _STEAP1; NP001162459.1_ Anubis babo (P. anubis) _STEAP1; NP_999470.1_ pig (S. scrofa) _STEAP1; And NP_081675.2_Mouse (M.musculus) _STEAP1.

NP_005663.2_ Human _PSCA is the registration number of the PSCA amino acid sequence available from GenBank.

                         SEQUENCE LISTING <110> THE TRUSTEES OF THE UNIVERSITY OF PENNSYLVANIA        WEINER, David        YAN, Jian        FERRARO, Bernadette        SARDESAI, Niranjan        RAMANATHAN, Mathura   <120> CONSENSUS PROSTATE ANTIGENS, NUCLEIC ACID MOLECULE ENCODING THE        SAME AND VACCINE AND USES COMPRISING THE SAME <130> UPVG0036 &Lt; 150 > US 61 / 413,176 <151> 2010-11-12 &Lt; 150 > US 61 / 417,817 <151> 2010-11-29 <160> 32 <170> PatentIn version 3.5 <210> 1 <211> 789 <212> DNA <213> Artificial Sequence <220> <223> PSA antigen 1 nucleic acid consensus sequence <400> 1 atgtgggtcc tggtggtgtt cctgactctg agcgtcacat ggatcggcgc cgctccactg 60 attctgagcc gcctggtggg cgggtgggag tgcgaaaagc actcccagcc atggcaggtg 120 ctggtcgctt ctaggggccg agcagtgtgc ggaggcgtgc tggtccaccc tcagtgggtc 180 ctgaccgcag cccattgtat ccgacagaag agcgtgattc tgctggggcg acaccagcca 240 ttctaccccg aggacacagg acaggtgttc caggtctctc acagttttcc ccatcctctg 300 tacaacatga gcctgctgaa aaacagatat ctgggacctg gcgacgatag ctcccatgat 360 ctgatgctgc tgaggctgtc cgagccagcc gaactgactg acgctgtgca ggtcctggat 420 ctgcccaccc aggagcctgc cctgggaacc acatgttatg cttcaggctg ggggagcatc 480 gaaccagagg aacatctgac tcccaagaaa ctgcagtgcg tggacctgca cctgattagt 540 aacgatgtgt gtgcacaggt ccattcacag aaggtgacaa agttcatgct gtgcgccggc 600 tcttggatgg gcggcaagtc aacttgcagc ggggactccg gcgggccact ggtgtgtgat 660 ggagtcctgc agggcatcac ctcttggggc agtcagcctt gtgccctgcc tcggagacca 720 agtctgtaca ctaaggtggt ccggtatagg aaatggattc aggacactat tgccgctaac 780 ccctgataa 789 <210> 2 <211> 261 <212> PRT <213> Artificial Sequence <220> <223> PSA antigen 1 amino acid consensus sequence <400> 2 Met Trp Val Leu Val Val Phe Leu Thr Leu Ser Val Thr Trp Ile Gly 1 5 10 15 Ala Ala Pro Leu Ile Leu Ser Arg Leu Val Gly Gly Trp Glu Cys Glu             20 25 30 Lys His Ser Gln Pro Trp Gln Val Leu Val Ala Ser Arg Gly Arg Ala         35 40 45 Val Cys Gly Gly Val Leu Val His Pro Gln Trp Val Leu Thr Ala Ala     50 55 60 His Cys Ile Arg Gln Lys Ser Val Ile Leu Leu Gly Arg His Gln Pro 65 70 75 80 Phe Tyr Pro Glu Asp Thr Gly Gln Val Phe Gln Val Ser His Ser Phe                 85 90 95 Pro His Pro Leu Tyr Asn Met Ser Leu Leu Lys Asn Arg Tyr Leu Gly             100 105 110 Pro Gly Asp Asp Ser Ser His Asp Leu Met Leu Leu Arg Leu Ser Glu         115 120 125 Pro Ala Glu Leu Thr Asp Ala Val Gln Val Leu Asp Leu Pro Thr Gln     130 135 140 Glu Pro Ala Leu Gly Thr Thr Cys Tyr Ala Ser Gly Trp Gly Ser Ile 145 150 155 160 Glu Pro Glu Glu His Leu Thr Pro Lys Lys Leu Gln Cys Val Asp Leu                 165 170 175 His Leu Ile Ser Asn Asp Val Cys Ala Gln Val His Ser Gln Lys Val             180 185 190 Thr Lys Phe Met Leu Cys Ala Gly Ser Trp Met Gly Gly Lys Ser Thr         195 200 205 Cys Ser Gly Asp Ser Gly Gly Pro Leu Val Cys Asp Gly Val Leu Gln     210 215 220 Gly Ile Thr Ser Trp Gly Ser Gln Pro Cys Ala Leu Pro Arg Arg Pro 225 230 235 240 Ser Leu Tyr Thr Lys Val Val Arg Tyr Arg Lys Trp Ile Gln Asp Thr                 245 250 255 Ile Ala Ala Asn Pro             260 <210> 3 <211> 840 <212> DNA <213> Artificial Sequence <220> <223> PSA antigen 2 nucleic acid consensus sequence <400> 3 atggactgga catggattct gttcctggtc gccgccgcaa ctcgcgtgca ttcctgggtc 60 ctggtggtgt tcctgactct gagcgtcaca tggatcggcg ccgctccact gattctgagc 120 cgcctggtgg gcgggtggga gtgcgaaaag cactcccagc catggcaggt gctggtcgct 180 tctaggggcc gagcagtgtg cggaggcgtg ctggtccacc ctcagtgggt cctgaccgca 240 gcccattgta tccgacagaa gagcgtgatt ctgctggggc gacaccagcc attctacccc 300 gaggacacag gacaggtgtt ccaggtctct cacagttttc cccatcctct gtacaacatg 360 agcctgctga aaaacagata tctgggacct ggcgacgata gctcccatga tctgatgctg 420 ctgaggctgt ccgagccagc cgaactgact gacgctgtgc aggtcctgga tctgcccacc 480 caggagcctg ccctgggaac cacatgttat gcttcaggct gggggagcat cgaaccagag 540 gaacatctga ctcccaagaa actgcagtgc gtggacctgc acctgattag taacgatgtg 600 tgtgcacagg tccattcaca gaaggtgaca aagttcatgc tgtgcgccgg ctcttggatg 660 ggcggcaagt caacttgcag cggggactcc ggcgggccac tggtgtgtga tggagtcctg 720 cagggcatca cctcttgggg cagtcagcct tgtgccctgc ctcggagacc aagtctgtac 780 actaaggtgg tccggtatag gaaatggatt caggacacta ttgccgctaa cccctgataa 840 <210> 4 <211> 278 <212> PRT <213> Artificial Sequence <220> <223> PSA antigen 2 amino acid consensus sequence <400> 4 Met Asp Trp Thr Trp Ile Leu Phe Leu Val Ala Ala Ala Thr Arg Val 1 5 10 15 His Ser Trp Val Leu Val Val Phe Leu Thr Leu Ser Val Thr Trp Ile             20 25 30 Gly Ala Pro Leu Ile Leu Ser Arg Leu Val Gly Gly Trp Glu Cys         35 40 45 Glu Lys His Ser Gln Pro Trp Gln Val Leu Val Ala Ser Arg Gly Arg     50 55 60 Ala Val Cys Gly Gly Val Leu Val His Pro Gln Trp Val Leu Thr Ala 65 70 75 80 Ala His Cys Ile Arg Gln Lys Ser Val Ile Leu Leu Gly Arg His Gln                 85 90 95 Pro Phe Tyr Pro Glu Asp Thr Gly Gln Val Phe Gln Val Ser His Ser             100 105 110 Phe Pro His Pro Leu Tyr Asn Met Ser Leu Leu Lys Asn Arg Tyr Leu         115 120 125 Gly Pro Gly Asp Asp Ser Ser His Asp Leu Met Leu Leu Arg Leu Ser     130 135 140 Glu Pro Ala Glu Leu Thr Asp Ala Val Gln Val Leu Asp Leu Pro Thr 145 150 155 160 Gln Glu Pro Ala Leu Gly Thr Thr Cys Tyr Ala Ser Gly Trp Gly Ser                 165 170 175 Ile Glu Pro Glu Glu His Leu Thr Pro Lys Lys Leu Gln Cys Val Asp             180 185 190 Leu His Leu Ile Ser Asn Asp Val Cys Ala Gln Val His Ser Gln Lys         195 200 205 Val Thr Lys Phe Met Leu Cys Ala Gly Ser Trp Met Gly Gly Lys Ser     210 215 220 Thr Cys Ser Gly Asp Ser Gly Gly Pro Leu Val Cys Asp Gly Val Leu 225 230 235 240 Gln Gly Ile Thr Ser Trp Gly Ser Gln Pro Cys Ala Leu Pro Arg Arg                 245 250 255 Pro Ser Leu Tyr Thr Lys Val Val Arg Tyr Arg Lys Trp Ile Gln Asp             260 265 270 Thr Ile Ala Ala Asn Pro         275 <210> 5 <211> 2283 <212> DNA <213> Artificial Sequence <220> <223> PSMA antigen 1 nucleic acid consensus sequence <400> 5 atgtggaacg cactgcatga gactgattct gctgtcgcac tgggacggag accccggtgg 60 ctgtgcgctg gagcactggt gctggccggc gggggattcc tgctgggatt cctgtttggc 120 tggtttatca aaagctccag cgaggctacc aatattaccc ctaagcacaa taagaaagca 180 ttcctggatg aactgaaagc cgagaacatc aagaaattcc tgtacaactt cacaagaatt 240 ccacatctgg ctggcactga gcagaacttc cagctggcaa aacagatcca gagtcagtgg 300 aaggaatttg ggctggactc agtggagctg acccactacg atgtcctgct gtcctatcca 360 aataagactc atcccaacta catctctatc attaacgaag acggaaatga gattttcaac 420 acctctctgt ttgaaccccc tccacccggc tatgagaatg tcagtgacgt ggtccctcca 480 ttctcagcct tcagccccca ggggatgcct gagggagatc tggtgtacgt caattatgct 540 agaacagaag acttctttaa gctggagagg gatatgaaaa tcaactgttc cggcaagatc 600 gtgattgccc ggtacgggaa ggtgttcaga ggaaataagg tcaaaaacgc tcagctggcc 660 ggagctaccg gcgtgatcct gtacagcgac cccgctgatt attttgcacc tggcgtgaag 720 tcctatccag acggatggaa tctgcccggc gggggagtgc agaggggaaa catcctgaac 780 ctgaatggag ccggcgatcc tctgactcca ggataccccg ccaacgaata cgcttatcgc 840 cggggaattg cagaggccgt gggcctgcct agcatcccag tccatcccat tggctattac 900 gatgcccaga agctgctgga gaaaatgggc gggagcgctc cccctgactc tagttggaag 960 ggctccctga aagtgcctta caatgtcggg ccaggattca ctgggaactt ttctacccag 1020 aaggtgaaaa tgcacatcca tagtaccagc gaggtgacac gaatctacaa cgtcattggc 1080 accctgagag gcgccgtgga gcctgatcgc tatgtcattc tgggaggcca cagagactca 1140 tgggtgttcg ggggaatcga tccacagagc ggagcagctg tggtccatga aattgtgcgc 1200 agctttggga ccctgaagaa agagggatgg cgacccaggc gcacaatcct gttcgcatcc 1260 tgggacgccg aggaatttgg gctgctgggc agcacagaat gggccgagga aaattctcgc 1320 ctgctgcagg agcgaggggt ggcttacatc aatgcagact caagcattga aggaaactat 1380 accctgcggg tggattgcac acccctgatg tacagtctgg tctataacct gacaaaggag 1440 ctgaaatcac ctgacgaggg cttcgaaggg aaaagcctgt acgaatcctg gactgagaag 1500 agcccatccc ccgaattcag cggcatgcct aggatctcta agctgggcag tgggaacgat 1560 tttgaggtgt tctttcagcg cctgggaatt gcctctggcc gagctcggta cacaaaaaat 1620 tgggagacta acaagttctc ctcttaccca ctgtatcaca gcgtgtacga gacttatgaa 1680 ctggtcgaga aattctacga ccccactttt aagtatcatc tgaccgtggc acaggtcagg 1740 ggcgggatgg tgttcgaact ggccaatagc atcgtcctgc catttgactg tcgagattac 1800 gctgtggtcc tgcggaagta cgcagacaag atctataaca tctccatgaa gcacccccag 1860 gagatgaagg cctattctgt gagtttcgat tccctgtttt ctgccgtcaa aaatttcacc 1920 gaaatcgcta gtaagttttc agagcgcctg caggacctgg ataagtccaa tcccatcctg 1980 ctgcggatta tgaacgatca gctgatgttc ctggaaagag cctttatcga ccctctgggc 2040 ctgcctgata gaccattcta caggcacgtg atctacgcac ctagttcaca taacaagtac 2100 gccggcgagt ctttcccagg gatctatgac gctctgtttg atattgaatc aaaggtggac 2160 cccagcaaag catggggcga ggtcaagaga cagatcagca ttgcagcctt tacagtgcag 2220 gccgccgccg aaaccctgtc cgaagtcgct tacccatacg atgtccccga ttacgcatga 2280 taa 2283 <210> 6 <211> 750 <212> PRT <213> Artificial Sequence <220> <223> PSMA antigen 1 amino acid consensus sequence <400> 6 Met Trp Asn Ala Leu His Glu Thr Asp Ser Ala Val Ala Leu Gly Arg 1 5 10 15 Arg Pro Arg Trp Leu Cys Ala Gly Ala Leu Val Leu Ala Gly Gly Gly             20 25 30 Phe Leu Leu Gly Phe Leu Phe Gly Trp Phe Ile Lys Ser Ser Ser Glu         35 40 45 Ala Thr Asn Ile Thr Pro Lys His Asn Lys Lys Ala Phe Leu Asp Glu     50 55 60 Leu Lys Ala Glu Asn Ile Lys Lys Phe Leu Tyr Asn Phe Thr Arg Ile 65 70 75 80 Pro His Leu Ala Gly Thr Glu Gln Asn Phe Gln Leu Ala Lys Gln Ile                 85 90 95 Gln Ser Gln Trp Lys Glu Phe Gly Leu Asp Ser Val Glu Leu Thr His             100 105 110 Tyr Asp Val Leu Leu Ser Tyr Pro Asn Lys Thr His Pro Asn Tyr Ile         115 120 125 Ser Ile Ile Asn Glu Asp Gly Asn Glu Ile Phe Asn Thr Ser Leu Phe     130 135 140 Glu Pro Pro Pro Gly Tyr Glu Asn Val Ser Asp Val Val Pro Pro 145 150 155 160 Phe Ser Ala Phe Ser Pro Gln Gly Met Pro Glu Gly Asp Leu Val Tyr                 165 170 175 Val Asn Tyr Ala Arg Thr Glu Asp Phe Phe Lys Leu Glu Arg Asp Met             180 185 190 Lys Ile Asn Cys Ser Gly Lys Ile Val Ile Ala Arg Tyr Gly Lys Val         195 200 205 Phe Arg Gly Asn Lys Val Lys Asn Ala Gln Leu Ala Gly Ala Thr Gly     210 215 220 Val Ile Leu Tyr Ser Asp Pro Ala Asp Tyr Phe Ala Pro Gly Val Lys 225 230 235 240 Ser Tyr Pro Asp Gly Trp Asn Leu Pro Gly Gly Gly Val Gln Arg Gly                 245 250 255 Asn Ile Leu Asn Leu Asn Gly Ala Gly Asp Pro Leu Thr Pro Gly Tyr             260 265 270 Pro Ala Asn Glu Tyr Ala Tyr Arg Arg Gly Ile Ala Glu Ala Val Gly         275 280 285 Leu Pro Ser Ile Pro Val His Pro Ile Gly Tyr Tyr Asp Ala Gln Lys     290 295 300 Leu Leu Glu Lys Met Gly Gly Ser Ala Pro Pro Asp Ser Ser Trp Lys 305 310 315 320 Gly Ser Leu Lys Val Pro Tyr Asn Val Gly Pro Gly Phe Thr Gly Asn                 325 330 335 Phe Ser Thr Gln Lys Val Lys Met His Ile His Ser Thr Ser Glu Val             340 345 350 Thr Arg Ile Tyr Asn Val Ile Gly Thr Leu Arg Gly Ala Val Glu Pro         355 360 365 Asp Arg Tyr Val Ile Leu Gly Gly His Arg Asp Ser Trp Val Phe Gly     370 375 380 Gly Ile Asp Pro Gln Ser Gly Ala Ala Val Val His Glu Ile Val Arg 385 390 395 400 Ser Phe Gly Thr Leu Lys Lys Glu Gly Trp Arg Pro Arg Arg Thr Ile                 405 410 415 Leu Phe Ala Ser Trp Asp Ala Glu Glu Phe Gly Leu Leu Gly Ser Thr             420 425 430 Glu Trp Ala Glu Glu Asn Ser Arg Leu Leu Gln Glu Arg Gly Val Ala         435 440 445 Tyr Ile Asn Ala Asp Ser Ser Ile Glu Gly Asn Tyr Thr Leu Arg Val     450 455 460 Asp Cys Thr Pro Leu Met Tyr Ser Leu Val Tyr Asn Leu Thr Lys Glu 465 470 475 480 Leu Lys Ser Pro Asp Glu Gly Phe Glu Gly Lys Ser Leu Tyr Glu Ser                 485 490 495 Trp Thr Glu Lys Ser Pro Ser Pro Glu Phe Ser Gly Met Pro Arg Ile             500 505 510 Ser Lys Leu Gly Ser Gly Asn Asp Phe Glu Val Phe Phe Gln Arg Leu         515 520 525 Gly Ile Ala Ser Gly Arg Ala Arg Tyr Thr Lys Asn Trp Glu Thr Asn     530 535 540 Lys Phe Ser Ser Tyr Pro Leu Tyr His Ser Val Tyr Glu Thr Tyr Glu 545 550 555 560 Leu Val Glu Lys Phe Tyr Asp Pro Thr Phe Lys Tyr His Leu Thr Val                 565 570 575 Ala Gln Val Arg Gly Gly Met Val Phe Glu Leu Ala Asn Ser Ile Val             580 585 590 Leu Pro Phe Asp Cys Arg Asp Tyr Ala Val Val Leu Arg Lys Tyr Ala         595 600 605 Asp Lys Ile Tyr Asn Ile Ser Met Lys His Pro Gln Glu Met Lys Ala     610 615 620 Tyr Ser Val Ser Phe Asp Ser Leu Phe Ser Ala Val Lys Asn Phe Thr 625 630 635 640 Glu Ile Ala Ser Lys Phe Ser Glu Arg Leu Gln Asp Leu Asp Lys Ser                 645 650 655 Asn Pro Ile Leu Leu Arg Ile Met Asn Asp Gln Leu Met Phe Leu Glu             660 665 670 Arg Ala Phe Ile Asp Pro Leu Gly Leu Pro Asp Arg Pro Phe Tyr Arg         675 680 685 His Val Ile Tyr Ala Pro Ser Ser His His Lys Tyr Ala Gly Glu Ser     690 695 700 Phe Pro Gly Ile Tyr Asp Ala Leu Phe Asp Ile Glu Ser Lys Val Asp 705 710 715 720 Pro Ser Lys Ala Trp Gly Glu Val Lys Arg Gln Ile Ser Ile Ala Ala                 725 730 735 Phe Thr Val Gln Ala Ala Ala Glu Thr Leu Ser Glu Val Ala             740 745 750 <210> 7 <211> 2334 <212> DNA <213> Artificial Sequence <220> <223> PSMA antigen 2 nucleic acid consensus sequence <400> 7 atggactgga catggattct gttcctggtc gccgccgcaa ctcgcgtgca ttcctggaac 60 gcactgcatg agactgattc tgctgtcgca ctgggacgga gaccccggtg gctgtgcgct 120 ggagcactgg tgctggccgg cgggggattc ctgctgggat tcctgtttgg ctggtttatc 180 aaaagctcca gcgaggctac caatattacc cctaagcaca ataagaaagc attcctggat 240 gaactgaaag ccgagaacat caagaaattc ctgtacaact tcacaagaat tccacatctg 300 gctggcactg agcagaactt ccagctggca aaacagatcc agagtcagtg gaaggaattt 360 gggctggact cagtggagct gacccactac gatgtcctgc tgtcctatcc aaataagact 420 catcccaact acatctctat cattaacgaa gacggaaatg agattttcaa cacctctctg 480 tttgaacccc ctccacccgg ctatgagaat gtcagtgacg tggtccctcc attctcagcc 540 ttcagccccc aggggatgcc tgagggagat ctggtgtacg tcaattatgc tagaacagaa 600 gacttcttta agctggagag ggatatgaaa atcaactgtt ccggcaagat cgtgattgcc 660 cggtacggga aggtgttcag aggaaataag gtcaaaaacg ctcagctggc cggagctacc 720 ggcgtgatcc tgtacagcga ccccgctgat tattttgcac ctggcgtgaa gtcctatcca 780 gacggatgga atctgcccgg cgggggagtg cagaggggaa acatcctgaa cctgaatgga 840 gccggcgatc ctctgactcc aggatacccc gccaacgaat acgcttatcg ccggggaatt 900 gcagaggccg tgggcctgcc tagcatccca gtccatccca ttggctatta cgatgcccag 960 aagctgctgg agaaaatggg cgggagcgct ccccctgact ctagttggaa gggctccctg 1020 aaagtgcctt acaatgtcgg gccaggattc actgggaact tttctaccca gaaggtgaaa 1080 atgtaccag atagtaccag cgaggtgaca cgaatctaca acgtcattgg caccctgaga 1140 ggcgccgtgg agcctgatcg ctatgtcatt ctgggaggcc acagagactc atgggtgttc 1200 gggggaatcg atccacagag cggagcagct gtggtccatg aaattgtgcg cagctttggg 1260 accctgaaga aagagggatg gcgacccagg cgcacaatcc tgttcgcatc ctgggacgcc 1320 gaggaatttg ggctgctggg cagcacagaa tgggccgagg aaaattctcg cctgctgcag 1380 gagcgagggg tggcttacat caatgcagac tcaagcattg aaggaaacta taccctgcgg 1440 gtggattgca cacccctgat gtacagtctg gtctataacc tgacaaagga gctgaaatca 1500 cctgacgagg gcttcgaagg gaaaagcctg tacgaatcct ggactgagaa gagcccatcc 1560 cccgaattca gcggcatgcc taggatctct aagctgggca gtgggaacga ttttgaggtg 1620 ttctttcagc gcctgggaat tgcctctggc cgagctcggt acacaaaaaa ttgggagact 1680 aacaagttct cctcttaccc actgtatcac agcgtgtacg agacttatga actggtcgag 1740 aaattctacg accccacttt taagtatcat ctgaccgtgg cacaggtcag gggcgggatg 1800 gtgttcgaac tggccaatag catcgtcctg ccatttgact gtcgagatta cgctgtggtc 1860 ctgcggaagt acgcagacaa gatctataac atctccatga agcaccccca ggagatgaag 1920 gcctattctg tgagtttcga ttccctgttt tctgccgtca aaaatttcac cgaaatcgct 1980 agtaagtttt cagagcgcct gcaggacctg gataagtcca atcccatcct gctgcggatt 2040 atgaacgatc agctgatgtt cctggaaaga gcctttatcg accctctggg cctgcctgat 2100 agaccattct acaggcacgt gatctacgca cctagttcac ataacaagta cgccggcgag 2160 tctttcccag ggatctatga cgctctgttt gatattgaat caaaggtgga ccccagcaaa 2220 gcatggggcg aggtcaagag acagatcagc attgcagcct ttacagtgca ggccgccgcc 2280 gaaaccctgt ccgaagtcgc ttacccatac gatgtccccg attacgcatg ataa 2334 <210> 8 <211> 766 <212> PRT <213> Artificial Sequence <220> <223> PSMA antigen 2 amino acid consensus sequence <400> 8 Met Trp Thr Trp Ile Leu Phe Leu Val Ala Ala Ala Thr Arg Val His 1 5 10 15 Ser Trp Asn Ala Leu His Glu Thr Asp Ser Ala Val Ala Leu Gly Arg             20 25 30 Arg Pro Arg Trp Leu Cys Ala Gly Ala Leu Val Leu Ala Gly Gly Gly         35 40 45 Phe Leu Leu Gly Phe Leu Phe Gly Trp Phe Ile Lys Ser Ser Ser Glu     50 55 60 Ala Thr Asn Ile Thr Pro Lys His Asn Lys Lys Ala Phe Leu Asp Glu 65 70 75 80 Leu Lys Ala Glu Asn Ile Lys Lys Phe Leu Tyr Asn Phe Thr Arg Ile                 85 90 95 Pro His Leu Ala Gly Thr Glu Gln Asn Phe Gln Leu Ala Lys Gln Ile             100 105 110 Gln Ser Gln Trp Lys Glu Phe Gly Leu Asp Ser Val Glu Leu Thr His         115 120 125 Tyr Asp Val Leu Leu Ser Tyr Pro Asn Lys Thr His Pro Asn Tyr Ile     130 135 140 Ser Ile Ile Asn Glu Asp Gly Asn Glu Ile Phe Asn Thr Ser Leu Phe 145 150 155 160 Glu Pro Pro Pro Gly Tyr Glu Asn Val Ser Asp Val Val Pro Pro                 165 170 175 Phe Ser Ala Phe Ser Pro Gln Gly Met Pro Glu Gly Asp Leu Val Tyr             180 185 190 Val Asn Tyr Ala Arg Thr Glu Asp Phe Phe Lys Leu Glu Arg Asp Met         195 200 205 Lys Ile Asn Cys Ser Gly Lys Ile Val Ile Ala Arg Tyr Gly Lys Val     210 215 220 Phe Arg Gly Asn Lys Val Lys Asn Ala Gln Leu Ala Gly Ala Thr Gly 225 230 235 240 Val Ile Leu Tyr Ser Asp Pro Ala Asp Tyr Phe Ala Pro Gly Val Lys                 245 250 255 Ser Tyr Pro Asp Gly Trp Asn Leu Pro Gly Gly Gly Val Gln Arg Gly             260 265 270 Asn Ile Leu Asn Leu Asn Gly Ala Gly Asp Pro Leu Thr Pro Gly Tyr         275 280 285 Pro Ala Asn Glu Tyr Ala Tyr Arg Arg Gly Ile Ala Glu Ala Val Gly     290 295 300 Leu Pro Ser Ile Pro Val His Pro Ile Gly Tyr Tyr Asp Ala Gln Lys 305 310 315 320 Leu Leu Glu Lys Met Gly Gly Ser Ala Pro Pro Asp Ser Ser Trp Lys                 325 330 335 Gly Ser Leu Lys Val Pro Tyr Asn Val Gly Pro Gly Phe Thr Gly Asn             340 345 350 Phe Ser Thr Gln Lys Val Lys Met His Ile His Ser Thr Ser Glu Val         355 360 365 Thr Arg Ile Tyr Asn Val Ile Gly Thr Leu Arg Gly Ala Val Glu Pro     370 375 380 Asp Arg Tyr Val Ile Leu Gly Gly His Arg Asp Ser Trp Val Phe Gly 385 390 395 400 Gly Ile Asp Pro Gln Ser Gly Ala Ala Val Val His Glu Ile Val Arg                 405 410 415 Ser Phe Gly Thr Leu Lys Lys Glu Gly Trp Arg Pro Arg Arg Thr Ile             420 425 430 Leu Phe Ala Ser Trp Asp Ala Glu Glu Phe Gly Leu Leu Gly Ser Thr         435 440 445 Glu Trp Ala Glu Glu Asn Ser Arg Leu Leu Gln Glu Arg Gly Val Ala     450 455 460 Tyr Ile Asn Ala Asp Ser Ser Ile Glu Gly Asn Tyr Thr Leu Arg Val 465 470 475 480 Asp Cys Thr Pro Leu Met Tyr Ser Leu Val Tyr Asn Leu Thr Lys Glu                 485 490 495 Leu Lys Ser Pro Asp Glu Gly Phe Glu Gly Lys Ser Leu Tyr Glu Ser             500 505 510 Trp Thr Glu Lys Ser Pro Ser Pro Glu Phe Ser Gly Met Pro Arg Ile         515 520 525 Ser Lys Leu Gly Ser Gly Asn Asp Phe Glu Val Phe Phe Gln Arg Leu     530 535 540 Gly Ile Ala Ser Gly Arg Ala Arg Tyr Thr Lys Asn Trp Glu Thr Asn 545 550 555 560 Lys Phe Ser Ser Tyr Pro Leu Tyr His Ser Val Tyr Glu Thr Tyr Glu                 565 570 575 Leu Val Glu Lys Phe Tyr Asp Pro Thr Phe Lys Tyr His Leu Thr Val             580 585 590 Ala Gln Val Arg Gly Gly Met Val Phe Glu Leu Ala Asn Ser Ile Val         595 600 605 Leu Pro Phe Asp Cys Arg Asp Tyr Ala Val Val Leu Arg Lys Tyr Ala     610 615 620 Asp Lys Ile Tyr Asn Ile Ser Met Lys His Pro Gln Glu Met Lys Ala 625 630 635 640 Tyr Ser Val Ser Phe Asp Ser Leu Phe Ser Ala Val Lys Asn Phe Thr                 645 650 655 Glu Ile Ala Ser Lys Phe Ser Glu Arg Leu Gln Asp Leu Asp Lys Ser             660 665 670 Asn Pro Ile Leu Leu Arg Ile Met Asn Asp Gln Leu Met Phe Leu Glu         675 680 685 Arg Ala Phe Ile Asp Pro Leu Gly Leu Pro Asp Arg Pro Phe Tyr Arg     690 695 700 His Val Ile Tyr Ala Pro Ser Ser His His Lys Tyr Ala Gly Glu Ser 705 710 715 720 Phe Pro Gly Ile Tyr Asp Ala Leu Phe Asp Ile Glu Ser Lys Val Asp                 725 730 735 Pro Ser Lys Ala Trp Gly Glu Val Lys Arg Gln Ile Ser Ile Ala Ala             740 745 750 Phe Thr Val Gln Ala Ala Ala Glu Thr Leu Ser Glu Val Ala         755 760 765 <210> 9 <211> 1023 <212> DNA <213> Artificial Sequence <220> <223> STEAP antigen 1 nucleic acid consensus sequence <400> 9 atggagagcc gcaaggacat cacaaatcag gaagagctgt ggaagatgaa accacggaga 60 aacctggagg aagacgatta cctgcacaag gacaccggcg aaacaagtat gctgaaaaga 120 ccagtgctgc tgcacctgca tcagactgct catgcagacg agtttgattg cccctctgaa 180 ctgcagcaca cccaggagct gttcccacag tggcatctgc ccatcaagat tgccgctatc 240 attgcttcac tgacatttct gtatactctg ctgagagaag tgatccaccc tctggccacc 300 agccatcagc agtacttcta taagatccct attctggtca tcaacaaggt cctgccaatg 360 gtgagcatca cactgctggc cctggtctac ctgcctggcg tgatcgcagc cattgtccag 420 ctgcacaacg gaacaaagta caagaagttc ccacattggc tggataagtg gatgctgact 480 aggaaacagt tcgggctgct gtccttcttt ttcgccgtgc tgcacgctat ctacagcctg 540 tcctatccca tgaggcgctc ttaccgatat aagctgctga actgggctta ccagcaggtg 600 cagcagaaca aggaggacgc atggattgaa cacgatgtgt ggcggatgga aatctatgtg 660 tctctgggca ttgtcgggct ggccatcctg gctctgctgg cagtgaccag tatcccttct 720 gtcagtgact cactgacatg gcgcgagttt cactacattc agagcaagct gggaatcgtg 780 tccctgctgc tgggcaccat ccatgcactg atttttgcct ggaataagtg gatcgatatc 840 aagcagttcg tgtggtatac tccccctacc tttatgattg ccgtcttcct gcccatcgtg 900 gtcctgattt ttaagtccat cctgttcctg ccttgtctgc gaaagaaaat cctgaaaatc 960 cgacatgggt gggaagacgt gacaaaaatc aataagaccg aaatctcaag ccagctgtga 1020 taa 1023 <210> 10 <211> 339 <212> PRT <213> Artificial Sequence <220> <223> STEAP antigen 1 amino acid consensus sequence <400> 10 Met Glu Ser Arg Lys Asp Ile Thr Asn Gln Glu Glu Leu Trp Lys Met 1 5 10 15 Lys Pro Arg Arg Asn Leu Glu Glu Asp Asp Tyr Leu His Lys Asp Thr             20 25 30 Gly Glu Thr Ser Met Leu Lys Arg Pro Val Leu Leu His Leu His Gln         35 40 45 Thr Ala His Ala Asp Glu Phe Asp Cys Pro Ser Glu Leu Gln His Thr     50 55 60 Gln Glu Leu Phe Pro Gln Trp His Leu Pro Ile Lys Ile Ala Ala Ile 65 70 75 80 Ile Ala Ser Leu Thr Phe Leu Tyr Thr Leu Leu Arg Glu Val Ile His                 85 90 95 Pro Leu Ala Thr Ser His Gln Gln Tyr Phe Tyr Lys Ile Pro Ile Leu             100 105 110 Val Ile Asn Lys Val Leu Pro Met Val Ser I Le Thu Leu Leu Ala Leu         115 120 125 Val Tyr Leu Pro Gly Val Ile Ala Ile Val Gln Leu His Asn Gly     130 135 140 Thr Lys Tyr Lys Lys Phe Pro His Trp Leu Asp Lys Trp Met Leu Thr 145 150 155 160 Arg Lys Gln Phe Gly Leu Leu Ser Phe Phe Phe Ala Val Leu His Ala                 165 170 175 Ile Tyr Ser Leu Ser Tyr Pro Met Arg Arg Ser Tyr Arg Tyr Lys Leu             180 185 190 Leu Asn Trp Ala Tyr Gln Gln Val Gln Gln Asn Lys Glu Asp Ala Trp         195 200 205 Ile Glu His Asp Val Trp Arg Met Glu Ile Tyr Val Ser Leu Gly Ile     210 215 220 Val Gly Leu Ala Ile Leu Ala Leu Leu Ala Val Thr Ser Ile Pro Ser 225 230 235 240 Val Ser Asp Ser Leu Thr Trp Arg Glu Phe His Tyr Ile Gln Ser Lys                 245 250 255 Leu Gly Ile Val Ser Leu Leu Leu Gly Thr Ile His Ala Leu Ile Phe             260 265 270 Ala Trp Asn Lys Trp Ile Asp Ile Lys Gln Phe Val Trp Tyr Thr Pro         275 280 285 Pro Thr Phe Met Ile Ala Val Phe Leu Pro Ile Val Val Leu Ile Phe     290 295 300 Lys Ser Ile Leu Phe Leu Pro Cys Leu Arg Lys Lys Ile Leu Lys Ile 305 310 315 320 Arg His Gly Trp Glu Asp Val Thr Lys Ile Asn Lys Thr Glu Ile Ser                 325 330 335 Ser Gln Leu              <210> 11 <211> 1074 <212> DNA <213> Artificial Sequence <220> <223> STEAP antigen 2 nucleic acid consensus sequence <400> 11 atggactgga catggattct gtttctggtc gctgccgcaa cccgcgtgca ttcagagagc 60 cgcaaggaca tcacaaatca ggaagagctg tggaagatga aaccacggag aaacctggag 120 gaagacgatt acctgcacaa ggacaccggc gaaacaagta tgctgaaaag accagtgctg 180 ctgcacctgc atcagactgc tcatgcagac gagtttgatt gcccctctga actgcagcac 240 acccaggagc tgttcccaca gtggcatctg cccatcaaga ttgccgctat cattgcttca 300 ctgacatttc tgtatactct gctgagagaa gtgatccacc ctctggccac cagccatcag 360 cagtacttct ataagatccc tattctggtc atcaacaagg tcctgccaat ggtgagcatc 420 acactgctgg ccctggtcta cctgcctggc gtgatcgcag ccattgtcca gctgcacaac 480 ggaacaaagt acaagaagtt cccacattgg ctggataagt ggatgctgac taggaaacag 540 ttcgggctgc tgtccttctt tttcgccgtg ctgcacgcta tctacagcct gtcctatccc 600 atgaggcgct cttaccgata taagctgctg aactgggctt accagcaggt gcagcagaac 660 aaggaggacg catggattga acacgatgtg tggcggatgg aaatctatgt gtctctgggc 720 attgtcgggc tggccatcct ggctctgctg gcagtgacca gtatcccttc tgtcagtgac 780 tcactgacat ggcgcgagtt tcactacatt cagagcaagc tgggaatcgt gtccctgctg 840 ctgggcacca tccatgcact gatttttgcc tggaataagt ggatcgatat caagcagttc 900 gtgtggtata ctccccctac ctttatgatt gccgtcttcc tgcccatcgt ggtcctgatt 960 tttaagtcca tcctgttcct gccttgtctg cgaaagaaaa tcctgaaaat ccgacatggg 1020 tgggaagacg tgacaaaaat caataagacc gaaatctcaa gccagctgtg ataa 1074 <210> 12 <211> 356 <212> PRT <213> Artificial Sequence <220> <223> STEAP antigen 2 amino acid consensus sequence <400> 12 Met Asp Trp Thr Trp Ile Leu Phe Leu Val Ala Ala Ala Thr Arg Val 1 5 10 15 His Ser Glu Ser Arg Lys Asp Ile Thr Asn Gln Glu Glu Leu Trp Lys             20 25 30 Met Lys Pro Arg Arg Asn Leu Glu Glu Asp Asp Tyr Leu His Lys Asp         35 40 45 Thr Gly Glu Thr Ser Met Leu Lys Arg Pro Val Leu Leu His Leu His     50 55 60 Gln Thr Ala His Ala Asp Glu Phe Asp Cys Pro Ser Glu Leu Gln His 65 70 75 80 Thr Gln Glu Leu Phe Pro Gln Trp His Leu Pro Ile Lys Ile Ala Ala                 85 90 95 Ile Ile Ala Ser Leu Thr Phe Leu Tyr Thr Leu Leu Arg Glu Val Ile             100 105 110 His Pro Leu Ala Thr Ser His Gln Gln Tyr Phe Tyr Lys Ile Pro Ile         115 120 125 Leu Val Ile Asn Lys Val Leu Pro Met Val Ser Ile Thr Leu Leu Ala     130 135 140 Leu Val Tyr Leu Pro Gly Val Ile Ala Ala Val Val Gln Leu His Asn 145 150 155 160 Gly Thr Lys Tyr Lys Lys Phe Pro His Trp Leu Asp Lys Trp Met Leu                 165 170 175 Thr Arg Lys Gln Phe Gly Leu Leu Ser Phe Phe Phe Ala Val Leu His             180 185 190 Ala Ile Tyr Ser Leu Ser Tyr Pro Met Arg Arg Ser Tyr Arg Tyr Lys         195 200 205 Leu Leu Asn Trp Ala Tyr Gln Gln Val Glu Gln Asn Lys Glu Asp Ala     210 215 220 Trp Ile Glu His Asp Val Trp Arg Met Glu Ile Tyr Val Ser Leu Gly 225 230 235 240 Ile Val Gly Leu Ile Leu Ile Leu Leu Ile Val Thr Ser Ile Pro                 245 250 255 Ser Val Ser Asp Ser Leu Thr Trp Arg Glu Phe His Tyr Ile Gln Ser             260 265 270 Lys Leu Gly Ile Val Ser Leu Leu Leu Gly Thr Ile His Ala Leu Ile         275 280 285 Phe Ala Trp Asn Lys Trp Ile Asp Ile Lys Gln Phe Val Trp Tyr Thr     290 295 300 Pro Pro Thr Phe Met Ile Ala Val Phe Leu Pro Ile Val Val Leu Ile 305 310 315 320 Phe Lys Ser Ile Leu Phe Leu Pro Cys Leu Arg Lys Lys Ile Leu Lys                 325 330 335 Ile Arg His Gly Trp Glu Asp Val Thr Lys Ile Asn Lys Thr Glu Ile             340 345 350 Ser Ser Gln Leu         355 <210> 13 <211> 399 <212> DNA <213> Artificial Sequence <220> &Lt; 223 > PSCA antigen nucleic acid consensus sequence <400> 13 atggactgga catggattct gtttctggtc gccgccgcaa cccgcgtgca ttctgctggc 60 ctggcactgc agcctggaac cgccctgctg tgctactctt gtaaggccca ggtgagtaac 120 gaggactgcc tgcaggtcga aaattgtact cagctgggag agcagtgctg gaccgcacgg 180 atcagagcag tgggactgct gacagtcatt agcaaagggt gctccctgaa ctgtgtggac 240 gatagccagg attactatgt cggaaagaaa aacatcacct gctgtgacac agatctgtgt 300 aatgcttctg gcgcccacgc tctgcagccc gcagccgcta ttctggctct gctgcccgct 360 ctgggactgc tgctgtgggg acccggacag ctgtgataa 399 <210> 14 <211> 131 <212> PRT <213> Artificial Sequence <220> <223> PSCA antigen amino acid consensus sequence <400> 14 Met Asp Trp Thr Trp Ile Leu Phe Leu Val Ala Ala Ala Thr Arg Val 1 5 10 15 His Ser Ala Gly Leu Ala Leu Gln Pro Gly Thr Ala Leu Leu Cys Tyr             20 25 30 Ser Cys Lys Ala Gln Val Ser Asn Glu Asp Cys Leu Gln Val Glu Asn         35 40 45 Cys Thr Gln Leu Gly Glu Gln Cys Trp Thr Ala Arg Ile Arg Ala Val     50 55 60 Gly Leu Leu Thr Val Ile Ser Lys Gly Cys Ser Leu Asn Cys Val Asp 65 70 75 80 Asp Ser Gln Asp Tyr Tyr Val Gly Lys Lys Asn Ile Thr Cys Cys Asp                 85 90 95 Thr Asp Leu Cys Asn Ala Ser Gly Ala His Ala Leu Gln Pro Ala Ala             100 105 110 Ala Ile Leu Ala Leu Leu Pro Ala Leu Gly Leu Leu Leu Trp Gly Pro         115 120 125 Gly Gln Leu     130 <210> 15 <211> 54 <212> DNA <213> Artificial Sequence <220> <223> IgE leader nucleic acid sequence <400> 15 atggactgga catggattct gtttctggtc gctgccgcaa cccgcgtgca ttca 54 <210> 16 <211> 17 <212> PRT <213> Artificial Sequence <220> <223> IgE leader amino acid sequence <400> 16 Met Trp Thr Trp Ile Leu Phe Leu Val Ala Ala Ala Thr Arg Val His 1 5 10 15 Ser      <210> 17 <211> 261 <212> PRT <213> Homo sapiens <400> 17 Met Trp Val Val Val Phe Leu Thr Leu Ser Val Thr Trp Ile Gly 1 5 10 15 Ala Ala Pro Leu Ile Leu Ser Arg Ile Val Gly Gly Trp Glu Cys Glu             20 25 30 Lys His Ser Gln Pro Trp Gln Val Leu Val Ala Ser Arg Gly Arg Ala         35 40 45 Val Cys Gly Gly Val Leu Val His Pro Gln Trp Val Leu Thr Ala Ala     50 55 60 His Cys Ile Arg Asn Lys Ser Val Ile Leu Leu Gly Arg His Ser Leu 65 70 75 80 Phe His Pro Glu Asp Thr Gly Gln Val Phe Gln Val Ser His Ser Phe                 85 90 95 Pro His Pro Leu Tyr Asp Met Ser Leu Leu Lys Asn Arg Phe Leu Arg             100 105 110 Pro Gly Asp Asp Ser Ser His Asp Leu Met Leu Leu Arg Leu Ser Glu         115 120 125 Pro Ala Glu Leu Thr Asp Ala Val Lys Val Met Asp Leu Pro Thr Gln     130 135 140 Glu Pro Ala Leu Gly Thr Thr Cys Tyr Ala Ser Gly Trp Gly Ser Ile 145 150 155 160 Glu Pro Glu Glu Phe Leu Thr Pro Lys Lys Leu Gln Cys Val Asp Leu                 165 170 175 His Val Ile Ser Asn Asp Val Cys Ala Gln Val His Pro Gln Lys Val             180 185 190 Thr Lys Phe Met Leu Cys Ala Gly Arg Trp Thr Gly Gly Lys Ser Thr         195 200 205 Cys Ser Gly Asp Ser Gly Gly Pro Leu Val Cys Asn Gly Val Leu Gln     210 215 220 Gly Ile Thr Ser Trp Gly Ser Glu Pro Cys Ala Leu Pro Glu Arg Pro 225 230 235 240 Ser Leu Tyr Thr Lys Val Val His Tyr Arg Lys Trp Ile Lys Asp Thr                 245 250 255 Ile Val Ala Asn Pro             260 <210> 18 <211> 262 <212> PRT <213> Homo sapiens <400> 18 Met Trp Val Val Val Phe Leu Thr Leu Ser Val Thr Trp Ile Gly 1 5 10 15 Ala Ala Pro Leu Ile Leu Ser Arg Ile Val Gly Gly Trp Glu Cys Glu             20 25 30 Lys His Ser Gln Pro Trp Gln Val Leu Val Ala Ser Arg Gly Arg Ala         35 40 45 Val Cys Gly Gly Val Leu Val His Pro Gln Trp Val Leu Thr Ala Ala     50 55 60 His Cys Ile Arg Lys Cys Lys Ser Val Ile Leu Leu Gly Arg His Ser 65 70 75 80 Leu Phe His Pro Glu Asp Thr Gly Gln Val Phe Gln Val Ser His Ser                 85 90 95 Phe Pro His Pro Leu Tyr Asp Met Ser Leu Leu Lys Asn Arg Phe Leu             100 105 110 Arg Pro Gly Asp Asp Ser Ser His Asp Leu Met Leu Leu Arg Leu Ser         115 120 125 Glu Pro Ala Glu Leu Thr Asp Ala Val Lys Val Met Asp Leu Pro Thr     130 135 140 Gln Glu Pro Ala Leu Gly Thr Thr Cys Tyr Ala Ser Gly Trp Gly Ser 145 150 155 160 Ile Glu Pro Glu Glu Phe Leu Thr Pro Lys Lys Leu Gln Cys Val Asp                 165 170 175 Leu His Val Ile Ser Asn Asp Val Cys Ala Gln Val His Pro Gln Lys             180 185 190 Val Thr Lys Phe Met Leu Cys Ala Gly Arg Trp Thr Gly Gly Lys Ser         195 200 205 Thr Cys Ser Gly Asp Ser Gly Gly Pro Leu Val Cys Asn Gly Val Leu     210 215 220 Gln Gly Ile Thr Ser Trp Gly Ser Glu Pro Cys Ala Leu Pro Glu Arg 225 230 235 240 Pro Ser Leu Tyr Thr Lys Val Val His Tyr Arg Lys Trp Ile Lys Asp                 245 250 255 Thr Ile Val Ala Asn Pro             260 <210> 19 <211> 261 <212> PRT <213> Macaca fascicularis <400> 19 Met Trp Val Leu Val Val Phe Leu Thr Leu Ser Val Thr Trp Ile Gly 1 5 10 15 Ala Ala Pro Leu Ile Leu Ser Arg Ile Val Gly Gly Trp Glu Cys Glu             20 25 30 Lys His Ser Gln Pro Trp Gln Val Leu Val Ala Ser His Gly Arg Ala         35 40 45 Val Cys Gly Gly Val Leu Val His Pro Gln Trp Val Leu Thr Ala Ala     50 55 60 His Cys Ile Arg Ser His Ser Val Ile Leu Leu Gly Arg His Asn Pro 65 70 75 80 Tyr Tyr Pro Glu Asp Thr Gly Gln Val Phe Gln Val Ser His Ser Phe                 85 90 95 Pro His Pro Leu Tyr Asn Met Ser Leu Leu Lys Asn Arg Tyr Leu Gly             100 105 110 Pro Gly Asp Asp Ser Ser His Asp Leu Met Leu Leu Arg Leu Ser Glu         115 120 125 Pro Ala Glu Ile Thr Asp Ala Val Gln Val Leu Asp Leu Pro Thr Trp     130 135 140 Glu Pro Glu Leu Gly Thr Thr Cys Tyr Ala Ser Gly Trp Gly Ser Ile 145 150 155 160 Glu Pro Glu Glu His Leu Thr Pro Lys Lys Leu Gln Cys Val Asp Leu                 165 170 175 His Ile Ile Ser Asn Asp Val Cys Ala Gln Val His Ser Gln Lys Val             180 185 190 Thr Lys Phe Met Leu Cys Ala Gly Ser Trp Met Gly Gly Lys Ser Thr         195 200 205 Cys Ser Gly Asp Ser Gly Gly Pro Leu Val Cys Asp Gly Val Leu Gln     210 215 220 Gly Ile Thr Ser Trp Gly Ser Gln Pro Cys Ala Leu Pro Arg Arg Pro 225 230 235 240 Ser Leu Tyr Thr Lys Val Val Arg Tyr Arg Lys Trp Ile Gln Asp Thr                 245 250 255 Ile Met Ala Asn Pro             260 <210> 20 <211> 261 <212> PRT <213> Macaca mulatta <400> 20 Met Trp Val Leu Val Val Phe Leu Thr Leu Ser Val Thr Trp Ile Gly 1 5 10 15 Ala Ala Pro Leu Ile Leu Ser Arg Ile Val Gly Gly Trp Glu Cys Glu             20 25 30 Lys His Ser Gln Pro Trp Gln Val Leu Val Ala Ser Arg Gly Arg Ala         35 40 45 Val Cys Gly Gly Val Leu Val His Pro Gln Trp Val Leu Thr Ala Ala     50 55 60 His Cys Ile Arg Ser Asn Ser Val Ile Leu Leu Gly Arg His Asn Pro 65 70 75 80 Tyr Tyr Pro Glu Asp Thr Gly Gln Val Phe Gln Val Ser His Ser Phe                 85 90 95 Pro His Pro Leu Tyr Asn Met Ser Leu Leu Lys Asn Arg Tyr Leu Gly             100 105 110 Pro Gly Asp Asp Ser Ser His Asp Leu Met Leu Leu Arg Leu Ser Glu         115 120 125 Pro Ala Glu Ile Thr Asp Ala Val Gln Val Leu Asp Leu Pro Thr Trp     130 135 140 Glu Pro Glu Leu Gly Thr Thr Cys Tyr Ala Ser Gly Trp Gly Ser Ile 145 150 155 160 Glu Pro Glu Glu His Leu Thr Pro Lys Lys Leu Gln Cys Val Asp Leu                 165 170 175 His Ile Ile Ser Asn Asp Val Cys Ala Gln Val His Ser Gln Lys Val             180 185 190 Thr Lys Phe Met Leu Cys Ala Gly Ser Trp Met Gly Gly Lys Ser Thr         195 200 205 Cys Ser Gly Asp Ser Gly Gly Pro Leu Val Cys Asp Gly Val Leu Gln     210 215 220 Gly Ile Thr Ser Trp Gly Ser Gln Pro Cys Ala Leu Pro Arg Arg Pro 225 230 235 240 Ser Leu Tyr Thr Lys Val Val Arg Tyr Arg Lys Trp Ile Gln Asp Thr                 245 250 255 Ile Met Ala Asn Pro             260 <210> 21 <211> 244 <212> PRT <213> Macaca mulatta <400> 21 Ala Pro Leu Ile Leu Ser Arg Ile Val Gly Gly Trp Glu Cys Glu Lys 1 5 10 15 His Ser Gln Pro Trp Gln Val Leu Val Ala Ser Arg Gly Arg Ala Val             20 25 30 Cys Gly Gly Val Leu Val His Pro Gln Trp Val Leu Thr Ala Ala His         35 40 45 Cys Ile Arg Ser Asn Ser Val Ile Leu Leu Gly Arg His Asn Pro Tyr     50 55 60 Tyr Pro Glu Asp Thr Gly Gln Val Phe Gln Val Ser Ser Ser Phe Pro 65 70 75 80 His Pro Leu Tyr Asn Met Ser Leu Leu Lys Asn Arg Tyr Leu Gly Pro                 85 90 95 Gly Asp As Ser Ser His Asp Leu Met Leu Leu Arg Leu Ser Glu Pro             100 105 110 Ala Glu Ile Thr Asp Ala Val Gln Val Leu Asp Leu Pro Thr Trp Glu         115 120 125 Pro Glu Leu Gly Thr Thr Cys Tyr Ala Ser Gly Trp Gly Ser Ile Glu     130 135 140 Pro Glu Glu His Leu Thr Pro Lys Lys Leu Gln Cys Val Asp Leu His 145 150 155 160 Ile Ile Ser Asn Asp Val Cys Ala Gln Val His Ser Gln Lys Val Thr                 165 170 175 Lys Phe Met Leu Cys Ala Gly Ser Trp Met Gly Gly Lys Ser Thr Cys             180 185 190 Ser Gly Asp Ser Gly Gly Pro Leu Val Cys Asp Gly Val Leu Gln Gly         195 200 205 Ile Thr Ser Trp Gly Ser Gln Pro Cys Ala Leu Pro Arg Arg Pro Ser     210 215 220 Leu Tyr Thr Lys Val Val Arg Tyr Arg Lys Trp Ile Gln Asp Thr Ile 225 230 235 240 Met Ala Asn Pro                  <210> 22 <211> 750 <212> PRT <213> Homo sapiens <400> 22 Met Trp Asn Leu Leu His Glu Thr Asp Ser Ala Val Ala Thr Ala Arg 1 5 10 15 Arg Pro Arg Trp Leu Cys Ala Gly Ala Leu Val Leu Ala Gly Gly Phe             20 25 30 Phe Leu Leu Gly Phe Leu Phe Gly Trp Phe Ile Lys Ser Ser Asn Glu         35 40 45 Ala Thr Asn Ile Thr Pro Lys His Asn Met Lys Ala Phe Leu Asp Glu     50 55 60 Leu Lys Ala Glu Asn Ile Lys Lys Phe Leu Tyr Asn Phe Thr Gln Ile 65 70 75 80 Pro His Leu Ala Gly Thr Glu Gln Asn Phe Gln Leu Ala Lys Gln Ile                 85 90 95 Gln Ser Gln Trp Lys Glu Phe Gly Leu Asp Ser Val Glu Leu Ala His             100 105 110 Tyr Asp Val Leu Leu Ser Tyr Pro Asn Lys Thr His Pro Asn Tyr Ile         115 120 125 Ser Ile Ile Asn Glu Asp Gly Asn Glu Ile Phe Asn Thr Ser Leu Phe     130 135 140 Glu Pro Pro Pro Gly Tyr Glu Asn Val Ser Asp Ile Val Pro Pro 145 150 155 160 Phe Ser Ala Phe Ser Pro Gln Gly Met Pro Glu Gly Asp Leu Val Tyr                 165 170 175 Val Asn Tyr Ala Arg Thr Glu Asp Phe Phe Lys Leu Glu Arg Asp Met             180 185 190 Lys Ile Asn Cys Ser Gly Lys Ile Val Ile Ala Arg Tyr Gly Lys Val         195 200 205 Phe Arg Gly Asn Lys Val Lys Asn Ala Gln Leu Ala Gly Ala Lys Gly     210 215 220 Val Ile Leu Tyr Ser Asp Pro Ala Asp Tyr Phe Ala Pro Gly Val Lys 225 230 235 240 Ser Tyr Pro Asp Gly Trp Asn Leu Pro Gly Gly Gly Val Gln Arg Gly                 245 250 255 Asn Ile Leu Asn Leu Asn Gly Ala Gly Asp Pro Leu Thr Pro Gly Tyr             260 265 270 Pro Ala Asn Glu Tyr Ala Tyr Arg Arg Gly Ile Ala Glu Ala Val Gly         275 280 285 Leu Pro Ser Ile Pro Val His Pro Ile Gly Tyr Tyr Asp Ala Gln Lys     290 295 300 Leu Leu Glu Lys Met Gly Gly Ser Ala Pro Pro Asp Ser Ser Trp Arg 305 310 315 320 Gly Ser Leu Lys Val Pro Tyr Asn Val Gly Pro Gly Phe Thr Gly Asn                 325 330 335 Phe Ser Thr Gln Lys Val Lys Met His Ile His Ser Thr Asn Glu Val             340 345 350 Thr Arg Ile Tyr Asn Val Ile Gly Thr Leu Arg Gly Ala Val Glu Pro         355 360 365 Asp Arg Tyr Val Ile Leu Gly Gly His Arg Asp Ser Trp Val Phe Gly     370 375 380 Gly Ile Asp Pro Gln Ser Gly Ala Ala Val Val His Glu Ile Val Arg 385 390 395 400 Ser Phe Gly Thr Leu Lys Lys Glu Gly Trp Arg Pro Arg Arg Thr Ile                 405 410 415 Leu Phe Ala Ser Trp Asp Ala Glu Glu Phe Gly Leu Leu Gly Ser Thr             420 425 430 Glu Trp Ala Glu Glu Asn Ser Arg Leu Leu Gln Glu Arg Gly Val Ala         435 440 445 Tyr Ile Asn Ala Asp Ser Ser Ile Glu Gly Asn Tyr Thr Leu Arg Val     450 455 460 Asp Cys Thr Pro Leu Met Tyr Ser Leu Val His Asn Leu Thr Lys Glu 465 470 475 480 Leu Lys Ser Pro Asp Glu Gly Phe Glu Gly Lys Ser Leu Tyr Glu Ser                 485 490 495 Trp Thr Lys Lys Ser Pro Ser Pro Glu Phe Ser Gly Met Pro Arg Ile             500 505 510 Ser Lys Leu Gly Ser Gly Asn Asp Phe Glu Val Phe Phe Gln Arg Leu         515 520 525 Gly Ile Ala Ser Gly Arg Ala Arg Tyr Thr Lys Asn Trp Glu Thr Asn     530 535 540 Lys Phe Ser Gly Tyr Pro Leu Tyr His Ser Val Tyr Glu Thr Tyr Glu 545 550 555 560 Leu Val Glu Lys Phe Tyr Asp Pro Met Phe Lys Tyr His Leu Thr Val                 565 570 575 Ala Gln Val Arg Gly Gly Met Val Phe Glu Leu Ala Asn Ser Ile Val             580 585 590 Leu Pro Phe Asp Cys Arg Asp Tyr Ala Val Val Leu Arg Lys Tyr Ala         595 600 605 Asp Lys Ile Tyr Ser Ile Ser Met Lys His Pro Gln Glu Met Lys Thr     610 615 620 Tyr Ser Val Ser Phe Asp Ser Leu Phe Ser Ala Val Lys Asn Phe Thr 625 630 635 640 Glu Ile Ala Ser Lys Phe Ser Glu Arg Leu Gln Asp Phe Asp Lys Ser                 645 650 655 Asn Pro Ile Val Leu Arg Met Met Asn Asp Gln Leu Met Phe Leu Glu             660 665 670 Arg Ala Phe Ile Asp Pro Leu Gly Leu Pro Asp Arg Pro Phe Tyr Arg         675 680 685 His Val Ile Tyr Ala Pro Ser Ser His His Lys Tyr Ala Gly Glu Ser     690 695 700 Phe Pro Gly Ile Tyr Asp Ala Leu Phe Asp Ile Glu Ser Lys Val Asp 705 710 715 720 Pro Ser Lys Ala Trp Gly Glu Val Lys Arg Gln Ile Tyr Val Ala Ala                 725 730 735 Phe Thr Val Gln Ala Ala Ala Glu Thr Leu Ser Glu Val Ala             740 745 750 <210> 23 <211> 749 <212> PRT <213> Homo sapiens <400> 23 Met Trp Asn Leu Leu His Glu Thr Asp Ser Ala Val Ala Thr Ala Arg 1 5 10 15 Arg Pro Arg Trp Leu Cys Ala Gly Ala Leu Val Leu Ala Gly Gly Phe             20 25 30 Phe Leu Leu Gly Phe Leu Phe Gly Trp Phe Ile Lys Ser Ser Asn Glu         35 40 45 Ala Thr Asn Ile Thr Pro Lys His Asn Met Lys Ala Phe Leu Asp Glu     50 55 60 Leu Lys Ala Glu Asn Ile Lys Lys Phe Leu His Asn Phe Thr Gln Ile 65 70 75 80 Pro His Leu Ala Gly Thr Glu Gln Asn Phe Gln Leu Ala Lys Gln Ile                 85 90 95 Gln Ser Gln Trp Lys Glu Phe Gly Leu Asp Ser Val Glu Leu Ala His             100 105 110 Tyr Asp Val Leu Leu Ser Tyr Pro Asn Lys Thr His Pro Asn Tyr Ile         115 120 125 Ser Ile Ile Asn Glu Asp Gly Asn Glu Ile Phe Asn Thr Ser Leu Phe     130 135 140 Glu Pro Pro Pro Gly Tyr Glu Asn Val Ser Asp Ile Val Pro Pro 145 150 155 160 Phe Ser Ala Phe Ser Pro Gln Gly Met Pro Glu Gly Asp Leu Val Tyr                 165 170 175 Val Asn Tyr Ala Arg Thr Glu Asp Phe Phe Lys Leu Glu Arg Asp Met             180 185 190 Lys Ile Asn Cys Ser Gly Lys Ile Val Ile Ala Arg Tyr Gly Lys Val         195 200 205 Phe Arg Gly Asn Lys Val Lys Asn Ala Gln Leu Ala Gly Ala Lys Gly     210 215 220 Val Ile Leu Tyr Ser Asp Pro Ala Asp Tyr Phe Ala Pro Gly Val Lys 225 230 235 240 Ser Tyr Pro Asp Gly Trp Asn Leu Pro Gly Gly Gly Val Gln Arg Gly                 245 250 255 Asn Ile Leu Asn Leu Asn Gly Ala Gly Asp Pro Leu Thr Pro Gly Tyr             260 265 270 Pro Ala Asn Glu Tyr Ala Tyr Arg Arg Gly Ile Ala Glu Ala Val Gly         275 280 285 Leu Pro Ser Ile Pro Val His Pro Ile Gly Tyr Tyr Asp Ala Gln Lys     290 295 300 Leu Leu Glu Lys Met Gly Gly Ser Ala Pro Pro Asp Ser Ser Trp Arg 305 310 315 320 Gly Ser Leu Lys Val Pro Tyr Asn Val Gly Pro Gly Phe Thr Gly Asn                 325 330 335 Phe Ser Thr Gln Lys Val Lys Met His Ile His Ser Thr Asn Glu Val             340 345 350 Thr Arg Ile Tyr Asn Val Ile Gly Thr Leu Arg Gly Ala Val Glu Pro         355 360 365 Asp Arg Tyr Val Ile Leu Gly Gly His Arg Asp Ser Trp Val Phe Gly     370 375 380 Gly Ile Asp Pro Gln Ser Gly Ala Ala Val Val His Glu Ile Val Arg 385 390 395 400 Ser Phe Gly Thr Leu Lys Lys Glu Gly Trp Arg Pro Arg Arg Thr Ile                 405 410 415 Leu Phe Ala Ser Trp Asp Ala Glu Glu Phe Gly Leu Leu Gly Ser Thr             420 425 430 Glu Trp Ala Glu Glu Asn Ser Arg Leu Leu Gln Glu Arg Gly Val Ala         435 440 445 Tyr Ile Asn Ala Asp Ser Ser Ile Glu Gly Asn Tyr Thr Leu Arg Val     450 455 460 Asp Cys Thr Pro Leu Met Tyr Ser Leu Val His Asn Leu Thr Lys Glu 465 470 475 480 Leu Lys Ser Pro Asp Glu Gly Phe Glu Gly Lys Ser Leu Tyr Glu Ser                 485 490 495 Trp Thr Lys Lys Ser Pro Ser Pro Glu Phe Ser Gly Met Pro Arg Ile             500 505 510 Ser Lys Leu Gly Ser Gly Asn Asp Phe Glu Val Phe Phe Gln Arg Leu         515 520 525 Gly Ile Ala Ser Gly Arg Ala Arg Tyr Thr Lys Asn Trp Glu Thr Asn     530 535 540 Phe Ser Gly Tyr Pro Leu Tyr His Ser Val Tyr Glu Thr Tyr Glu Leu 545 550 555 560 Val Glu Lys Phe Tyr Asp Pro Met Phe Lys Tyr His Leu Thr Val Ala                 565 570 575 Gln Val Arg Gly Gly Met Val Phe Glu Leu Ala Asn Ser Ile Val Leu             580 585 590 Pro Phe Asp Cys Arg Asp Tyr Ala Val Val Leu Arg Lys Tyr Ala Asp         595 600 605 Lys Ile Tyr Ser Ile Ser Met Lys His Pro Gln Glu Met Lys Thr Tyr     610 615 620 Ser Val Ser Phe Asp Ser Leu Phe Ser Ala Val Lys Asn Phe Thr Glu 625 630 635 640 Ile Ala Ser Lys Phe Ser Glu Arg Leu Gln Asp Phe Asp Lys Ser Asn                 645 650 655 Pro Ile Val Leu Arg Met Met Asn Asp Gln Leu Met Phe Leu Glu Arg             660 665 670 Ala Phe Ile Asp Pro Leu Gly Leu Pro Asp Arg Pro Phe Tyr Arg His         675 680 685 Val Ile Tyr Ala Pro Ser Ser His His Lys Tyr Ala Gly Glu Ser Phe     690 695 700 Pro Gly Ile Tyr Asp Ala Leu Phe Asp Ile Glu Ser Lys Val Asp Pro 705 710 715 720 Ser Lys Ala Trp Gly Glu Val Lys Arg Gln Ile Tyr Val Ala Ala Phe                 725 730 735 Thr Val Gln Ala Ala Ala Glu Thr Leu Ser Glu Val Ala             740 745 <210> 24 <211> 735 <212> PRT <213> Macaca mulatta <400> 24 Met Ile Ala Gly Ser Ser Tyr Pro Leu Leu Leu Ala Ala Tyr Ala Cys 1 5 10 15 Thr Gly Cys Leu Ala Glu Arg Leu Gly Trp Phe Ile Lys Ser Ser Ser             20 25 30 Glu Ala Thr Asn Ile Thr Pro Lys His Asn Met Lys Ala Phe Leu Asp         35 40 45 Glu Leu Lys Ala Glu Asn Ile Lys Lys Phe Leu His Asn Phe Thr Gln     50 55 60 Ile Pro His Leu Ala Gly Thr Glu Gln Asn Phe Gln Leu Ala Lys Gln 65 70 75 80 Ile Gln Ser Gln Trp Lys Glu Phe Gly Leu Asp Ser Val Glu Leu Thr                 85 90 95 His Tyr Asp Val Leu Leu Ser Tyr Pro Asn Lys Thr His Pro Asn Tyr             100 105 110 Ile Ser Ile Ile Asn Glu Asp Gly Asn Glu Ile Phe Asn Thr Ser Leu         115 120 125 Phe Glu Pro Pro Ala Gly Tyr Glu Asn Val Ser Asp Ile Val Pro     130 135 140 Pro Phe Ser Ala Phe Ser Pro Gln Gly Met Pro Glu Gly Asp Leu Val 145 150 155 160 Tyr Val Asn Tyr Ala Arg Thr Glu Asp Phe Phe Lys Leu Glu Arg Asp                 165 170 175 Met Lys Ile Asn Cys Ser Gly Lys Ile Val Ile Ala Arg Tyr Gly Lys             180 185 190 Val Phe Arg Gly Asn Lys Val Lys Asn Ala Gln Leu Ala Gly Ala Thr         195 200 205 Gly Val Ile Leu Tyr Ser Asp Pro Ala Asp Tyr Phe Ala Pro Gly Val     210 215 220 Lys Ser Tyr Pro Asp Gly Trp Asn Leu Pro Gly Gly Gly Val Gln Arg 225 230 235 240 Gly Asn Ile Leu Asn Leu Asn Gly Ala Gly Asp Pro Leu Thr Pro Gly                 245 250 255 Tyr Pro Ala Asn Glu Tyr Ala Tyr Arg Arg Gly Met Ala Glu Ala Val             260 265 270 Gly Leu Pro Ser Ile Pro Val His Pro Ile Gly Tyr Tyr Asp Ala Gln         275 280 285 Lys Leu Leu Glu Lys Met Gly Gly Ser Ala Ser Pro Asp Ser Ser Trp     290 295 300 Arg Gly Ser Leu Lys Val Pro Tyr Asn Val Gly Pro Gly Phe Thr Gly 305 310 315 320 Asn Phe Ser Thr Gln Lys Val Lys Met His Ile His Ser Thr Ser Glu                 325 330 335 Val Thr Arg Ile Tyr Asn Val Ile Gly Thr Leu Arg Gly Ala Val Glu             340 345 350 Pro Asp Arg Tyr Val Ile Leu Gly Gly His Arg Asp Ser Trp Val Phe         355 360 365 Gly Ile Asp Pro Gln Ser Gly Ala Ala Val Val His Glu Ile Val     370 375 380 Arg Ser Phe Gly Thr Leu Lys Lys Glu Gly Trp Arg Pro Arg Arg Thr 385 390 395 400 Ile Leu Phe Ala Ser Trp Asp Ala Glu Glu Phe Gly Leu Leu Gly Ser                 405 410 415 Thr Glu Trp Ala Glu Glu Asn Ser Arg Leu Leu Gln Glu Arg Gly Val             420 425 430 Ala Tyr Ile Asn Ala Asp Ser Ser Ile Glu Gly Asn Tyr Thr Leu Arg         435 440 445 Val Asp Cys Thr Pro Leu Met Tyr Ser Leu Val Tyr Asn Leu Thr Lys     450 455 460 Glu Leu Glu Ser Pro Asp Glu Gly Phe Glu Gly Lys Ser Leu Tyr Glu 465 470 475 480 Ser Trp Thr Lys Lys Ser Pro Ser Pro Glu Phe Ser Gly Met Pro Arg                 485 490 495 Ile Ser Lys Leu Gly Ser Gly Asn Asp Phe Glu Val Phe Phe Gln Arg             500 505 510 Leu Gly Ile Ala Ser Gly Arg Ala Arg Tyr Thr Lys Asn Trp Glu Thr         515 520 525 Asn Lys Phe Ser Ser Tyr Pro Leu Tyr His Ser Val Tyr Glu Thr Tyr     530 535 540 Glu Leu Val Glu Lys Phe Tyr Asp Pro Met Phe Lys Tyr His Leu Thr 545 550 555 560 Val Ala Gln Val Arg Gly Gly Met Val Phe Glu Leu Ala Asn Ser Val                 565 570 575 Val Leu Pro Phe Asp Cys Arg Asp Tyr Ala Val Val Leu Arg Lys Tyr             580 585 590 Ala Asp Lys Ile Tyr Asn Ile Ser Met Lys His Pro Gln Glu Met Lys         595 600 605 Thr Tyr Ser Val Ser Phe Asp Ser Leu Phe Ser Ala Val Lys Asn Phe     610 615 620 Thr Glu Ile Ala Ser Lys Phe Ser Glu Arg Leu Arg Asp Phe Asp Lys 625 630 635 640 Ser Asn Pro Ile Leu Leu Arg Met Met Asn Asp Gln Leu Met Phe Leu                 645 650 655 Glu Arg Ala Phe Ile Asp Pro Leu Gly Leu Pro Asp Arg Pro Phe Tyr             660 665 670 Arg His Val Ile Tyr Ala Pro Ser Ser His His Lys Tyr Ala Gly Glu         675 680 685 Ser Phe Pro Gly Ile Tyr Asp Ala Leu Phe Asp Ile Glu Ser Lys Val     690 695 700 Asp Pro Ser Gln Ala Trp Gly Glu Val Lys Arg Gln Ile Ser Ile Ala 705 710 715 720 Thr Phe Thr Val Gln Ala Ala Ala Glu Thr Leu Ser Glu Val Ala                 725 730 735 <210> 25 <211> 704 <212> PRT <213> Macaca mulatta <400> 25 Met Ile Ala Gly Ser Ser Tyr Pro Leu Leu Leu Ala Ala Tyr Ala Cys 1 5 10 15 Thr Gly Cys Leu Ala Glu Arg Leu Gly Trp Phe Ile Lys Ser Ser Ser             20 25 30 Glu Ala Thr Asn Ile Thr Pro Lys His Asn Met Lys Ala Phe Leu Asp         35 40 45 Glu Leu Lys Ala Glu Asn Ile Lys Lys Phe Leu His Asn Phe Thr Gln     50 55 60 Ile Pro His Leu Ala Gly Thr Glu Gln Asn Phe Gln Leu Ala Lys Gln 65 70 75 80 Ile Gln Ser Gln Trp Lys Glu Phe Gly Leu Asp Ser Val Glu Leu Thr                 85 90 95 His Tyr Asp Val Leu Leu Ser Tyr Pro Asn Lys Thr His Pro Asn Tyr             100 105 110 Ile Ser Ile Ile Asn Glu Asp Gly Asn Glu Ile Phe Asn Thr Ser Leu         115 120 125 Phe Glu Pro Pro Ala Gly Tyr Glu Asn Val Ser Asp Ile Val Pro     130 135 140 Pro Phe Ser Ala Phe Ser Pro Gln Gly Met Pro Glu Gly Asp Leu Val 145 150 155 160 Tyr Val Asn Tyr Ala Arg Thr Glu Asp Phe Phe Lys Leu Glu Arg Asp                 165 170 175 Met Lys Ile Asn Cys Ser Gly Lys Ile Val Ile Ala Arg Tyr Gly Lys             180 185 190 Val Phe Arg Gly Asn Lys Val Lys Asn Ala Gln Leu Ala Gly Ala Thr         195 200 205 Gly Val Ile Leu Tyr Ser Asp Pro Ala Asp Tyr Phe Ala Pro Gly Val     210 215 220 Lys Ser Tyr Pro Asp Gly Trp Asn Leu Pro Gly Gly Gly Val Gln Arg 225 230 235 240 Gly Asn Ile Leu Asn Leu Asn Gly Ala Gly Asp Pro Leu Thr Pro Gly                 245 250 255 Tyr Pro Ala Asn Glu Tyr Ala Tyr Arg Arg Gly Met Ala Glu Ala Val             260 265 270 Gly Leu Pro Ser Ile Pro Val His Pro Ile Gly Tyr Tyr Asp Ala Gln         275 280 285 Lys Leu Leu Glu Lys Met Gly Gly Ser Ala Ser Pro Asp Ser Ser Trp     290 295 300 Arg Gly Ser Leu Lys Val Pro Tyr Asn Val Gly Pro Gly Phe Thr Gly 305 310 315 320 Asn Phe Ser Thr Gln Lys Val Lys Met His Ile His Ser Thr Ser Glu                 325 330 335 Val Thr Arg Ile Tyr Asn Val Ile Gly Thr Leu Arg Gly Ala Val Glu             340 345 350 Pro Asp Arg Tyr Val Ile Leu Gly Gly His Arg Asp Ser Trp Val Phe         355 360 365 Gly Ile Asp Pro Gln Ser Gly Ala Ala Val Val His Glu Ile Val     370 375 380 Arg Ser Phe Gly Thr Leu Lys Lys Glu Gly Trp Arg Pro Arg Arg Thr 385 390 395 400 Ile Leu Phe Ala Ser Trp Asp Ala Glu Glu Phe Gly Leu Leu Gly Ser                 405 410 415 Thr Glu Trp Ala Glu Glu Asn Ser Arg Leu Leu Gln Glu Arg Gly Val             420 425 430 Ala Tyr Ile Asn Ala Asp Ser Ser Ile Glu Gly Asn Tyr Thr Leu Arg         435 440 445 Val Asp Cys Thr Pro Leu Met Tyr Ser Leu Val Tyr Asn Leu Thr Lys     450 455 460 Glu Leu Glu Ser Pro Asp Glu Gly Phe Glu Gly Lys Ser Leu Tyr Glu 465 470 475 480 Ser Trp Thr Lys Lys Ser Pro Ser Pro Glu Phe Ser Gly Met Pro Arg                 485 490 495 Ile Ser Lys Leu Gly Ser Gly Asn Asp Phe Glu Val Phe Phe Gln Arg             500 505 510 Leu Gly Ile Ala Ser Gly Arg Ala Arg Tyr Thr Lys Asn Trp Glu Thr         515 520 525 Asn Lys Phe Ser Ser Tyr Pro Leu Tyr His Ser Val Tyr Glu Thr Tyr     530 535 540 Glu Leu Val Glu Lys Phe Tyr Asp Pro Met Phe Lys Tyr His Leu Thr 545 550 555 560 Val Ala Gln Val Arg Gly Gly Met Val Phe Glu Leu Ala Asn Ser Val                 565 570 575 Val Leu Pro Phe Asp Cys Arg Asp Tyr Ala Val Val Leu Arg Lys Tyr             580 585 590 Ala Asp Lys Ile Tyr Asn Ile Ser Met Lys His Pro Gln Glu Met Lys         595 600 605 Thr Tyr Ser Val Ser Phe Asp Ser Leu Phe Ser Ala Val Lys Asn Phe     610 615 620 Thr Glu Ile Ala Ser Lys Phe Ser Glu Arg Leu Arg Asp Phe Asp Lys 625 630 635 640 Ser Lys His Val Ile Tyr Ala Pro Ser Ser His His Lys Tyr Ala Gly                 645 650 655 Glu Ser Phe Pro Gly Ile Tyr Asp Ala Leu Phe Asp Ile Glu Ser Lys             660 665 670 Val Asp Pro Ser Gln Ala Trp Gly Glu Val Lys Arg Gln Ile Ser Ile         675 680 685 Ala Thr Phe Thr Val Ala Ala Glu Thr Leu Ser Glu Val Ala     690 695 700 <210> 26 <211> 339 <212> PRT <213> Homo sapiens <400> 26 Met Glu Ser Arg Lys Asp Ile Thr Asn Gln Glu Glu Leu Trp Lys Met 1 5 10 15 Lys Pro Arg Arg Asn Leu Glu Glu Asp Asp Tyr Leu His Lys Asp Thr             20 25 30 Gly Glu Thr Ser Met Leu Lys Arg Pro Val Leu Leu His Leu His Gln         35 40 45 Thr Ala His Ala Asp Glu Phe Asp Cys Pro Ser Glu Leu Gln His Thr     50 55 60 Gln Glu Leu Phe Pro Gln Trp His Leu Pro Ile Lys Ile Ala Ala Ile 65 70 75 80 Ile Ala Ser Leu Thr Phe Leu Tyr Thr Leu Leu Arg Glu Val Ile His                 85 90 95 Pro Leu Ala Thr Ser His Gln Gln Tyr Phe Tyr Lys Ile Pro Ile Leu             100 105 110 Val Ile Asn Lys Val Leu Pro Met Val Ser I Le Thu Leu Leu Ala Leu         115 120 125 Val Tyr Leu Pro Gly Val Ile Ala Ile Val Gln Leu His Asn Gly     130 135 140 Thr Lys Tyr Lys Lys Phe Pro His Trp Leu Asp Lys Trp Met Leu Thr 145 150 155 160 Arg Lys Gln Phe Gly Leu Leu Ser Phe Phe Phe Ala Val Leu His Ala                 165 170 175 Ile Tyr Ser Leu Ser Tyr Pro Met Arg Arg Ser Tyr Arg Tyr Lys Leu             180 185 190 Leu Asn Trp Ala Tyr Gln Gln Val Gln Gln Asn Lys Glu Asp Ala Trp         195 200 205 Ile Glu His Asp Val Trp Arg Met Glu Ile Tyr Val Ser Leu Gly Ile     210 215 220 Val Gly Leu Ala Ile Leu Ala Leu Leu Ala Val Thr Ser Ile Pro Ser 225 230 235 240 Val Ser Asp Ser Leu Thr Trp Arg Glu Phe His Tyr Ile Gln Ser Lys                 245 250 255 Leu Gly Ile Val Ser Leu Leu Leu Gly Thr Ile His Ala Leu Ile Phe             260 265 270 Ala Trp Asn Lys Trp Ile Asp Ile Lys Gln Phe Val Trp Tyr Thr Pro         275 280 285 Pro Thr Phe Met Ile Ala Val Phe Leu Pro Ile Val Val Leu Ile Phe     290 295 300 Lys Ser Ile Leu Phe Leu Pro Cys Leu Arg Lys Lys Ile Leu Lys Ile 305 310 315 320 Arg His Gly Trp Glu Asp Val Thr Lys Ile Asn Lys Thr Glu Ile Cys                 325 330 335 Ser Gln Leu              <210> 27 <211> 339 <212> PRT <213> Homo sapiens <400> 27 Met Glu Ser Arg Lys Asp Ile Thr Asn Gln Glu Glu Leu Trp Lys Met 1 5 10 15 Lys Pro Arg Arg Asn Leu Glu Glu Asp Asp Tyr Leu His Lys Asp Thr             20 25 30 Gly Glu Thr Ser Met Leu Lys Arg Pro Val Leu Leu His Leu Gln Gln         35 40 45 Thr Ala His Ala Asp Glu Phe Asp Cys Pro Ser Glu Leu Gln His Thr     50 55 60 Gln Glu Leu Phe Pro Gln Trp His Leu Pro Ile Lys Ile Ala Ala Ile 65 70 75 80 Ile Ala Ser Leu Thr Phe Leu Tyr Thr Leu Leu Arg Glu Val Ile His                 85 90 95 Pro Leu Ala Thr Ser His Gln Gln Tyr Phe Tyr Lys Ile Pro Ile Leu             100 105 110 Val Ile Asn Lys Val Leu Pro Met Val Ser I Le Thu Leu Leu Ala Leu         115 120 125 Val Tyr Leu Pro Gly Val Ile Ala Ile Val Gln Leu His Asn Gly     130 135 140 Thr Lys Tyr Lys Lys Phe Pro His Trp Leu Asp Lys Trp Met Leu Thr 145 150 155 160 Arg Lys Gln Phe Gly Leu Leu Ser Phe Phe Phe Ala Val Leu His Ala                 165 170 175 Ile Tyr Ser Leu Ser Tyr Pro Met Arg Arg Ser Tyr Arg Tyr Lys Leu             180 185 190 Leu Asn Trp Ala Tyr Gln Gln Val Gln Gln Asn Lys Glu Asp Ala Trp         195 200 205 Ile Glu His Asp Val Trp Arg Met Glu Ile Tyr Val Ser Leu Gly Ile     210 215 220 Val Gly Leu Ala Ile Leu Ala Leu Leu Ala Val Thr Ser Ile Pro Ser 225 230 235 240 Val Ser Asp Ser Leu Thr Trp Arg Glu Phe His Tyr Ile Gln Ser Lys                 245 250 255 Leu Gly Ile Val Ser Leu Leu Leu Gly Thr Ile His Ala Leu Ile Phe             260 265 270 Ala Trp Asn Lys Trp Ile Asp Ile Lys Gln Phe Val Trp Tyr Thr Pro         275 280 285 Pro Thr Phe Met Ile Ala Val Phe Leu Pro Ile Val Val Leu Ile Phe     290 295 300 Lys Ser Ile Leu Phe Leu Pro Cys Leu Arg Lys Lys Ile Leu Lys Ile 305 310 315 320 Arg His Gly Trp Glu Asp Val Thr Lys Ile Asn Lys Thr Glu Ile Cys                 325 330 335 Ser Gln Leu              <210> 28 <211> 339 <212> PRT <213> Macaca mulatta <400> 28 Met Glu Ser Arg Lys Asp Ile Thr Asn Glu Glu Glu Leu Trp Lys Met 1 5 10 15 Lys Pro Arg Arg Asn Leu Glu Glu Asp Asp Tyr Leu His Lys Asp Thr             20 25 30 Gly Glu Thr Ser Met Leu Lys Arg Pro Val Leu Leu His Leu His Gln         35 40 45 Thr Ala His Ala Asp Glu Phe Asp Cys Pro Ser Glu Leu Gln His Thr     50 55 60 Gln Glu Leu Phe Pro Gln Trp His Leu Pro Ile Lys Ile Ala Ala Ile 65 70 75 80 Ile Ala Ser Leu Thr Phe Leu Tyr Thr Leu Leu Arg Glu Val Ile His                 85 90 95 Pro Leu Ala Thr Ser His Gln Gln Tyr Phe Tyr Lys Ile Pro Ile Leu             100 105 110 Val Ile Asn Lys Val Leu Pro Met Val Ser I Le Thu Leu Leu Ala Leu         115 120 125 Val Tyr Leu Pro Gly Val Ile Ala Ile Val Gln Leu His Asn Gly     130 135 140 Thr Lys Tyr Lys Lys Phe Pro His Trp Leu Asp Lys Trp Met Leu Thr 145 150 155 160 Arg Lys Gln Phe Gly Leu Leu Ser Phe Phe Phe Ala Val Leu His Ala                 165 170 175 Ile Tyr Ser Leu Ser Tyr Pro Met Arg Arg Ser Tyr Arg Tyr Lys Leu             180 185 190 Leu Asn Trp Ala Tyr Gln Gln Val Gln Gln Asn Lys Glu Asp Ala Trp         195 200 205 Ile Glu His Asp Val Trp Arg Met Glu Ile Tyr Val Ser Leu Gly Ile     210 215 220 Val Gly Leu Ala Ile Leu Ala Leu Leu Ala Val Thr Ser Ile Pro Ser 225 230 235 240 Val Ser Asp Ser Leu Thr Trp Arg Glu Phe His Tyr Ile Gln Ser Lys                 245 250 255 Leu Gly Ile Val Ser Leu Leu Ile Phe             260 265 270 Ala Trp Asn Lys Trp Ile Asp Ile Lys Gln Phe Val Trp Tyr Thr Pro         275 280 285 Pro Thr Phe Met Ile Ala Val Phe Leu Pro Val Val Val Leu Ile Phe     290 295 300 Lys Ser Ile Leu Phe Leu Pro Cys Leu Arg Lys Lys Ile Leu Lys Ile 305 310 315 320 Arg His Gly Trp Glu Asp Val Thr Lys Ile Asn Lys Met Glu Ile Ser                 325 330 335 Ser Gln Leu              <210> 29 <211> 259 <212> PRT <213> Homo sapiens <400> 29 Met Glu Ser Arg Lys Asp Ile Thr Asn Gln Glu Glu Ile Trp Lys Met 1 5 10 15 Lys Pro Arg Arg Asn Leu Glu Asp Asn Asp Tyr Leu His Lys Asp Thr             20 25 30 Gly Glu Thr Ser Met Leu Lys Arg Pro Val Leu Leu His Leu Gln Gln         35 40 45 Thr Ala His Ala Asp Glu Phe Asp Cys Pro Ser Glu Leu Gln His Ala     50 55 60 Gln Glu Leu Phe Pro Gln Trp His Leu Pro Ile Lys Ile Ala Ala Val 65 70 75 80 Met Ala Ser Leu Thr Phe Leu Tyr Thr Leu Leu Arg Glu Val Ile His                 85 90 95 Pro Leu Ala Thr Ser His Gln Gln Tyr Phe Tyr Lys Ile Pro Ile Leu             100 105 110 Val Ile Asn Lys Val Leu Pro Met Val Ser I Le Thu Leu Leu Ala Leu         115 120 125 Val Tyr Leu Pro Gly Val Ile Ala Ala Val Val Gln Val His Asn Gly     130 135 140 Thr Lys Tyr Lys Lys Phe Pro His Trp Leu Asp Lys Trp Met Leu Thr 145 150 155 160 Arg Lys Gln Phe Gly Leu Leu Ser Leu Phe Phe Ala Val Leu His Ala                 165 170 175 Ile Tyr Thr Leu Ser Tyr Ala Met Arg Arg Ser Tyr Arg Tyr Lys Leu             180 185 190 Leu Asn Trp Ala Tyr Gln Gln Val Gln Gln Asn Lys Glu Asp Ala Trp         195 200 205 Ile Glu His Asp Val Trp Arg Met Glu Ile Tyr Val Ser Leu Gly Ile     210 215 220 Val Gly Leu Ala Ile Leu Ala Leu Leu Ala Val Thr Ser Ile Pro Ser 225 230 235 240 Val Ser Asp Ser Leu Thr Trp Arg Glu Phe His Tyr Ile Gln Arg Leu                 245 250 255 Leu Gln Glu              <210> 30 <211> 258 <212> PRT <213> Homo sapiens <400> 30 Met Glu Ser Arg Lys Asp Ile Thr Asn Gln Glu Glu Ile Trp Lys Met 1 5 10 15 Lys Pro Arg Arg Asn Leu Glu Asp Asn Asp Tyr Leu His Lys Asp Thr             20 25 30 Gly Glu Thr Ser Met Leu Lys Arg Pro Val Leu Leu His Leu Gln Gln         35 40 45 Thr Ala His Ala Asp Glu Phe Asp Cys Pro Ser Glu Leu Gln His Ala     50 55 60 Gln Glu Leu Phe Pro Gln Trp His Leu Pro Ile Lys Ile Ala Ala Val 65 70 75 80 Met Ala Ser Leu Thr Phe Leu Tyr Thr Leu Leu Arg Glu Val Ile His                 85 90 95 Pro Leu Ala Thr Ser His Gln Gln Tyr Phe Tyr Lys Ile Pro Ile Leu             100 105 110 Val Ile Asn Lys Val Leu Pro Met Val Ser I Le Thu Leu Leu Ala Leu         115 120 125 Val Tyr Leu Pro Gly Val Ile Ala Ala Val Val Gln Val His Asn Gly     130 135 140 Thr Lys Tyr Lys Lys Phe Pro His Trp Leu Asp Lys Trp Met Leu Thr 145 150 155 160 Arg Lys Gln Phe Gly Leu Leu Ser Leu Phe Phe Ala Val Leu His Ala                 165 170 175 Ile Tyr Thr Leu Ser Tyr Ala Met Arg Arg Ser Tyr Arg Tyr Lys Leu             180 185 190 Leu Asn Trp Ala Tyr Gln Gln Val Gln Gln Asn Lys Glu Asp Ala Trp         195 200 205 Ile Glu His Asp Val Trp Arg Met Glu Ile Tyr Val Ser Leu Gly Ile     210 215 220 Val Gly Leu Ala Ile Leu Ala Leu Leu Ala Val Thr Ser Ile Pro Ser 225 230 235 240 Val Ser Asp Ser Leu Thr Trp Arg Glu Phe His Tyr Ile Gln Val Asn                 245 250 255 Asn Ile          <210> 31 <211> 114 <212> PRT <213> Homo sapiens <400> 31 Met Ala Gly Leu Ala Leu Gln Pro Gly Thr Ala Leu Leu Cys Tyr Ser 1 5 10 15 Cys Lys Ala Gln Val Ser Asn Glu Asp Cys Leu Gln Val Glu Asn Cys             20 25 30 Thr Gln Leu Gly Glu Gln Cys Trp Thr Ala Arg Ile Arg Ala Val Gly         35 40 45 Leu Leu Thr Val Ile Ser Lys Gly Cys Ser Leu Asn Cys Val Asp Asp     50 55 60 Ser Gln Asp Tyr Tyr Val Gly Lys Lys Asn Ile Thr Cys Cys Asp Thr 65 70 75 80 Asp Leu Cys Asn Ala Ser Gly Ala His Ala Leu Gln Pro Ala Ala Ala                 85 90 95 Ile Leu Ala Leu Leu Pro Ala Leu Gly Leu Leu Leu Trp Gly Pro Gly             100 105 110 Gln Leu <210> 32 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> HA Tag amino acid sequence <400> 32 Tyr Pro Tyr Asp Val Pro Asp Tyr Ala 1 5         

Claims (19)

A nucleic acid molecule comprising a coding sequence encoding at least one protein selected from the group consisting of:
a) SEQ ID NO: 2; A protein that is conserved and 98% homologous to SEQ ID NO: 2, wherein amino acids 69, 78, 80, 82, 102, 110, 137, 139, 165, 189, 203, 220, 232 and 248 of SEQ ID NO: Or amino acids 69, 78, 80, 82, 102, 110, 137, 139, 165, 189, 203, 220, 232 and 248 of SEQ ID NO: 2 are conserved and amino acids corresponding to at least 256 amino acid residues of SEQ ID NO: An immunogenic fragment of SEQ ID NO: 2; And
b) SEQ ID NO: 4; Amino acids 21, 86, 127, 129, 154, 156, 182, 195, 206, 218, 220, 237, 249, 255, 265, 271 and 275 of SEQ ID NO: 4 are conserved and 98% Adult protein; Or amino acids 21, 86, 127, 129, 154, 156, 182, 195, 206, 218, 220, 237, 249, 255, 265, 271 and 275 of SEQ ID NO: 4 are conserved and at least 274 amino acids 4. An immunogenic fragment of SEQ ID NO: 4 comprising an amino acid corresponding to a residue of SEQ ID NO: delete The nucleic acid molecule of claim 1, wherein the nucleic acid molecule further encodes one or more proteins selected from the group consisting of:
c) SEQ ID NO: 6; Amino acids 14, 15, 32, 47, 58, 79, 111, 157, 223, 320, 350, 475, 499, 569, 613, 624, 653, 660, 663, 733 and 734 of SEQ ID NO: A protein that is 98% homologous to SEQ ID NO: 6; Or amino acids 14, 15, 32, 47, 58, 79, 111, 157, 223, 320, 350, 475, 499, 569, 613, 624, 653, 660, 663, 733 and 734 of SEQ ID NO: An immunogenic fragment of SEQ ID NO: 6, comprising an amino acid corresponding to at least 735 amino acid residues of SEQ ID NO: 6; And
d) SEQ ID NO: 8; Amino acids 21, 31, 32, 49, 64, 75, 96, 128, 174, 240, 337, 367, 492, 516, 565, 586, 630, 641, 670, 677, 751 is conserved and is 98% homologous to SEQ ID NO: 8; Or amino acids 21, 31, 32, 49, 64, 75, 96, 128, 174, 240, 337, 367, 492, 516, 565, 586, 630, 641, 670, 677, 680, 750, And 751 are conserved and comprise an amino acid corresponding to at least the 751 amino acid residue of SEQ ID NO: 8. 4. The nucleic acid molecule of claim 1 or 3, wherein the nucleic acid molecule is selected from the group consisting of SEQ ID NO: 2; And SEQ ID &lt; RTI ID = 0.0 &gt; 4, &lt; / RTI &gt; delete The nucleic acid molecule according to claim 1, wherein the coding sequence is at least one selected from the group consisting of:
a) a coding sequence which is 98% homologous to SEQ ID NO: 1 or SEQ ID NO: 1; And
b) a coding sequence which is 98% homologous to SEQ ID NO: 3 or SEQ ID NO: 3. delete 7. The nucleic acid molecule according to claim 6, wherein the nucleic acid molecule further comprises at least one selected from the group consisting of:
c) a coding sequence which is 98% homologous to the nucleotides 1 to 2250 of SEQ ID NO: 5, or the nucleotides 1 to 2250 of SEQ ID NO: 5; And
d) a coding sequence which is 98% homologous to nucleotides 1-2301 of SEQ ID NO: 7, or nucleotides 1-2301 of SEQ ID NO: 7. 9. The nucleic acid molecule according to claim 6 or 8, wherein the nucleic acid molecule is selected from the group consisting of SEQ ID NO: 1; And SEQ ID NO: 3. 3. The nucleic acid molecule according to claim 1, wherein the nucleotide sequence is at least one nucleotide sequence selected from the group consisting of SEQ ID NO: 7. The nucleic acid molecule according to claim 1 or 6, wherein the nucleic acid molecule is a plasmid. 7. The nucleic acid molecule according to claim 1 or 6, wherein the nucleic acid molecule is an expression vector and the coding sequence is operatively linked to a regulatory element. A pharmaceutical composition for treating a subject diagnosed with prostate cancer, comprising the nucleic acid molecule of any one of claims 1 to 6. A composition for inducing an immune response to a prostate cancer antigen comprising a protein selected from the group consisting of:
a) SEQ ID NO: 2; A protein that is conserved and 98% homologous to SEQ ID NO: 2, wherein amino acids 69, 78, 80, 82, 102, 110, 137, 139, 165, 189, 203, 220, 232 and 248 of SEQ ID NO: Or amino acids 69, 78, 80, 82, 102, 110, 137, 139, 165, 189, 203, 220, 232 and 248 of SEQ ID NO: 2 are conserved and amino acids corresponding to at least 256 amino acid residues of SEQ ID NO: An immunogenic fragment of SEQ ID NO: 2; And
b) SEQ ID NO: 4; Amino acids 21, 86, 127, 129, 154, 156, 182, 195, 206, 218, 220, 237, 249, 255, 265, 271 and 275 of SEQ ID NO: 4 are conserved and 98% Adult protein; Or amino acids 21, 86, 127, 129, 154, 156, 182, 195, 206, 218, 220, 237, 249, 255, 265, 271 and 275 of SEQ ID NO: 4 are conserved and at least 274 amino acids 4. An immunogenic fragment of SEQ ID NO: 4 comprising an amino acid corresponding to a residue of SEQ ID NO: delete 14. The composition of claim 13, further comprising at least one protein selected from the group consisting of:
c) SEQ ID NO: 6; Amino acids 14, 15, 32, 47, 58, 79, 111, 157, 223, 320, 350, 475, 499, 569, 613, 624, 653, 660, 663, 733 and 734 of SEQ ID NO: A protein that is 98% homologous to SEQ ID NO: 6; Or amino acids 14, 15, 32, 47, 58, 79, 111, 157, 223, 320, 350, 475, 499, 569, 613, 624, 653, 660, 663, 733 and 734 of SEQ ID NO: An immunogenic fragment of SEQ ID NO: 6, comprising an amino acid corresponding to at least 735 amino acid residues of SEQ ID NO: 6; And
d) SEQ ID NO: 8; Amino acids 21, 31, 32, 49, 64, 75, 96, 128, 174, 240, 337, 367, 492, 516, 565, 586, 630, 641, 670, 677, 751 is conserved and is 98% homologous to SEQ ID NO: 8; Or amino acids 21, 31, 32, 49, 64, 75, 96, 128, 174, 240, 337, 367, 492, 516, 565, 586, 630, 641, 670, 677, 680, 750, And 751 are conserved and comprise an amino acid corresponding to at least the 751 amino acid residue of SEQ ID NO: 8. 16. The method according to claim 13 or 15, wherein SEQ ID NO: 2; And an amino acid sequence encoding a protein selected from the group consisting of SEQ ID NO: 4. delete 14. A pharmaceutical composition for treating an individual diagnosed with prostate cancer, comprising the composition of claim 13. 13. The pharmaceutical composition of claim 12, further comprising a pharmaceutically acceptable excipient.

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